Joel Goldstein

When exponentially growing Escherichia coli cell cultures were transferred from 37 degrees C to 10 degrees C or 15 degrees C, the production of a 7.4-kDa cytoplasmic protein (CS7.4) was prominently induced. The rate of CS7.4 production reached 13% of total protein synthesis within 1-1.5 hr after a shift to 10 degrees C and subsequently dropped to a lower basal level. Regulation of CS7.4 expression was very strict, such that synthesis of the protein was undetectable at 37 degrees C. We have cloned the gene encoding this protein and have completed the nucleotide sequence analysis, which revealed that the gene encodes a hydrophilic protein of 70 amino acid residues.

The design and construction of the silicon strip microvertex detector (SMD) of the L3 experiment at LEP are described. We present the sensors, readout electronics, data acquisition system, mechanical assembly and support, displacement monitoring systems and radiation monitoring system of the recently installed double-sided, double-layered SMD. This detector utilizes novel and sophisticated techniques for its readout.

A search for a standard model Higgs boson decaying into a pair of τ leptons is performed using events recorded by the CMS experiment at the LHC in 2011 and 2012. The dataset corresponds to an integrated luminosity of 4.9 fb−1 at a centre-of-mass energy of 7 TeV and 19.7 fb−1 at 8 TeV. Each τ lepton decays hadronically or leptonically to an electron or a muon, leading to six different final states for the τ -lepton pair, all considered in this analysis. An excess of events is observed over the expected background contributions, with a local significance larger than 3 standard deviations for m H values between 115 and 130 GeV. The best fit of the observed H → τ τ signal cross section times branching fraction for m H = 125 GeV is 0.78 ± 0.27 times the standard model expectation. These observations constitute evidence for the 125 GeV Higgs boson decaying to a pair of τ leptons.

Smallpox vaccination in the United States is a routine public health measure which has been under intensive review during the last decade. The most frequently occurring adverse reactions to vaccination are benign and require little or no systemic therapy. These reactions include accidental infection, erythematous and urticarial rash, and generalized vaccinia. Chickenpox occurring concurrently with vaccination presents no problem unless vaccinia has widely superinfected the chickenpox lesions. There is no risk to the pregnant woman who is vaccinated, but there is a slight risk that the fetus will develop fetal vaccinia. The vaccinia does not cause congenital malformations. Vaccinia hyperimmune globulin (VIG) in prophylactic dosage may be given to a pregnant woman who is traveling to a smallpox infected or endemic area in order to prevent fetal vaccinia. Vaccinia necrosum and eczema vaccinatum require vigorous systemic therapy with VIG, and often thiosemicarbazone. Post-vaccinial encephalitis, while frequently serious, has not been shown to be ameliorated by VIG therapy, although there are data which suggest VIG has some value in prophylaxis for encephalitis. Prophylaxis, prompt recognition, and proper therapy may reduce the fatality rates of these complications. Revaccination of patients who have suffered a complication is a frequent clinical problem. Revaccination of an individual who has had post-vaccinial encephalitis or vaccinia necrosum is contraindicated unless the risk of contracting smallpox outweighs the risk of the above two diseases. Revaccination of children who have had eczema vaccinatum is not contraindicated. Revaccination of children with a history of accidental infection or erythematous or urticarial rash presents no known or theoretically increased risk.

The Compact Linear Collider (CLIC) is an option for a future e+e- collider operating at centre-of-mass energies up to 3 TeV, providing sensitivity to a wide range of new physics phenomena and precision physics measurements at the energy frontier. This paper is the first comprehensive presentation of the Higgs physics reach of CLIC operating at three energy stages: sqrt(s) = 350 GeV, 1.4 TeV and 3 TeV. The initial stage of operation allows the study of Higgs boson production in Higgsstrahlung (e+e- -> ZH) and WW-fusion (e+e- -> Hnunu), resulting in precise measurements of the production cross sections, the Higgs total decay width Gamma_H, and model-independent determinations of the Higgs couplings. Operation at sqrt(s) > 1 TeV provides high-statistics samples of Higgs bosons produced through WW-fusion, enabling tight constraints on the Higgs boson couplings. Studies of the rarer processes e+e- -> ttH and e+e- -> HHnunu allow measurements of the top Yukawa coupling and the Higgs boson self-coupling. This paper presents detailed studies of the precision achievable with Higgs measurements at CLIC and describes the interpretation of these measurements in a global fit.

A search for neutral Higgs bosons in the minimal supersymmetric extension of the standard model (MSSM) decaying to tau-lepton pairs in pp collisions is performed, using events recorded by the CMS experiment at the LHC. The dataset corresponds to an integrated luminosity of 24.6 fb−1, with 4.9 fb−1 at 7 TeV and 19.7 fb−1 at 8 TeV. To enhance the sensitivity to neutral MSSM Higgs bosons, the search includes the case where the Higgs boson is produced in association with a b-quark jet. No excess is observed in the tau-lepton-pair invariant mass spectrum. Exclusion limits are presented in the MSSM parameter space for different benchmark scenarios, m h max , m h mod + , m h mod − , light-stop, light-stau, τ-phobic, and low-m H. Upper limits on the cross section times branching fraction for gluon fusion and b-quark associated Higgs boson production are also given.

A search is performed for long-lived massive neutral particles decaying to quark-antiquark pairs. The experimental signature is a distinctive topology of a pair of jets, originating at a secondary vertex. Events were collected with the CMS detector at the CERN LHC in proton-proton collisions at a centre-of-mass energy of 8 TeV. The data analyzed correspond to an integrated luminosity of 18.5 inverse femtobarns. No significant excess is observed above standard model expectations. Upper limits at 95% confidence level are set on the production cross section of a heavy neutral scalar particle, H, in the mass range of 200 to 1000 GeV, decaying promptly into a pair of long-lived neutral X particles in the mass range of 50 to 350 GeV, each in turn decaying into a quark-antiquark pair. For X with mean proper decay lengths of 0.4 to 200 cm, the upper limits are typically 0.5-200 fb. The results are also interpreted in the context of an R-parity-violating supersymmetric model with long-lived neutralinos decaying into a quark-antiquark pair and a muon. For pair production of squarks that promptly decay to neutralinos with mean proper decay lengths of 2-40 cm, the upper limits on the cross section are typically 0.5-3 fb. The above limits are the most stringent on these channels to date.

A search is performed for the production of heavy resonances decaying into top-antitop quark pairs in proton-proton collisions at sqrt(s) = 8 TeV. Data used for the analyses were collected with the CMS detector and correspond to an integrated luminosity of 19.7 inverse femtobarns. The search is performed using events with three different final states, defined by the number of leptons (electrons and muons) from the t t-bar to W b W b decay. The analyses are optimized for reconstruction of top quarks with high Lorentz boosts, where jet substructure techniques are used to enhance the sensitivity. Results are presented for all channels and a combination is performed. No significant excess of events relative to the expected yield from standard model processes is observed. Upper limits on the production cross section of heavy resonances decaying to t t-bar are calculated. A narrow leptophobic topcolor Z' resonance with a mass below 2.4 TeV is excluded at 95% confidence level. Limits are also derived for a broad Z' resonance with a 10% width relative to the resonance mass, and a Kaluza-Klein excitation of the gluon in the Randall-Sundrum model. These are the most stringent limits to date on heavy resonances decaying into top-antitop quark pairs.

The Mu3e experiment aims to find or exclude the lepton flavour violating decay μ→eee at branching fractions above 10−16. A first phase of the experiment using an existing beamline at the Paul Scherrer Institute (PSI) is designed to reach a single event sensitivity of 2⋅10−15. We present an overview of all aspects of the technical design and expected performance of the phase I Mu3e detector. The high rate of up to 108 muon decays per second and the low momenta of the decay electrons and positrons pose a unique set of challenges, which we tackle using an ultra thin tracking detector based on high-voltage monolithic active pixel sensors combined with scintillating fibres and tiles for precise timing measurements.

Abstract Background Mast cells (MC) are powerful inflammatory immune sentinel cells that drive numerous allergic, inflammatory, and pruritic disorders when activated. MC‐targeted therapies are approved in several disorders, yet many patients have limited benefit suggesting the need for approaches that more broadly inhibit MC activity. MCs require the KIT receptor and its ligand stem cell factor (SCF) for differentiation, maturation, and survival. Here we describe CDX‐0159, an anti‐KIT monoclonal antibody that potently suppresses MCs in human healthy volunteers. Methods CDX‐0159‐mediated KIT inhibition was tested in vitro using KIT‐expressing immortalized cells and primary human mast cells. CDX‐0159 safety and pharmacokinetics were evaluated in a 13‐week good laboratory practice (GLP)‐compliant cynomolgus macaque study. A single ascending dose (0.3, 1, 3, and 9 mg/kg), double‐blinded placebo‐controlled phase 1a human healthy volunteer study ( n = 32) was conducted to evaluate the safety, pharmacokinetics, and pharmacodynamics of CDX‐0159. Results CDX‐0159 inhibits SCF‐dependent KIT activation in vitro . Fc modifications in CDX‐0159 led to elimination of effector function and reduced serum clearance. In cynomolgus macaques, multiple high doses were safely administered without a significant impact on hematology, a potential concern for KIT inhibitors. A single dose of CDX‐0159 in healthy human subjects was generally well tolerated and demonstrated long antibody exposure. Importantly, CDX‐0159 led to dose‐dependent, profound suppression of plasma tryptase, a MC‐specific protease associated with tissue MC burden, indicative of systemic MC suppression or ablation. Conclusion CDX‐0159 administration leads to systemic mast cell ablation and may represent a safe and novel approach to treat mast cell‐driven disorders.

Epidermal growth factor receptor (EGF-R) overexpression is common in a large number of solid tumors and represents a negative prognostic indicator. Overexpression of EGF-R is strongly tumor associated, and this tyrosine kinase type receptor is considered an attractive target for Ab therapy. In this study, we describe the evaluation of mAb 2F8, a high avidity human mAb (IgG1kappa) directed against EGF-R, developed using human Ig transgenic mice. mAb 2F8 effectively blocked binding of EGF and TGF-alpha to the EGF-R. At saturating concentrations, 2F8 completely blocked EGF-R signaling and inhibited the in vitro proliferation of EGF-R-overexpressing A431 cells. At much lower concentrations, associated with low receptor occupancy, 2F8 induced efficient Ab-dependent cell-mediated cytotoxicity (ADCC) in vitro. In vivo studies showed potent antitumor effects in models with A431 tumor xenografts in athymic mice. Ex vivo analysis of the EGF-R status in tumor xenografts in 2F8-treated mice revealed that there are two therapeutic mechanisms. First, blocking of EGF-R signaling, which is most effective at complete receptor saturation and therefore requires a relatively high Ab dose. Second, at very low 2F8 receptor occupancy, we observed potent antitumor effects in mice, which are likely based on the engagement of immune effector mechanisms, in particular ADCC. Taken together, our findings indicate that ADCC represents an important effector mechanism of this Ab, which is effective at relatively low dose.

The Intermediate Silicon Layers (ISL) detector is currently being built as part of the CDF II detector upgrade project. The ISL detector will significantly improve tracking in the central region and, together with the Silicon Vertex detector, provide stand-alone 3D track information in the forward/backward regions. In this article, we present the quality of the production sensors manufactured by Hamamatsu Photonics, which account for roughly half of the silicon sensors used in the ISL detector.

The major cold shock protein of Escherichia coli, CS7.4, is produced at a level of 13% of total protein synthesis upon a temperature shift from 37 to 10 degrees C. The transcription of its gene (cspA) was found to be tightly regulated and induced only at low temperature. In addition, the cspA mRNA was extremely unstable at 37 degrees C, so that CS7.4 production was hardly detected when the culture temperature was shifted from 15 degrees C to 37 degrees C. The transcription initiation site (+1) was identified. In vivo footprinting demonstrated that the region from bases -35 to -73 was protected from chemical modification, and gel mobility shift analysis showed that a cold-shocked cell extract contained a factor(s) specifically bound to the fragment containing the sequence between bases -63 and -92. This factor was synthesized de novo only at low temperature, and its synthesis was inhibited by chloramphenicol. Possible functions of this factor are discussed.

Targeting recycling endocytic receptors with specific Abs provides a means for introducing a variety of tumor-associated Ags into human dendritic cells (DCs), culminating in their efficient presentation to T cells. We have generated a human mAb (B11) against the mannose receptor that is rapidly internalized by DCs through receptor-mediated endocytosis. By genetically linking the melanoma Ag, pmel17, to Ab B11, we obtained the fully human fusion protein, B11-pmel17. Treatment of DCs with B11-pmel17 resulted in the presentation of pmel17 in the context of HLA class I and class II molecules. Thus, potent pmel17-specific T cells were cytotoxic toward gp100(+) HLA-matched melanoma targets, but not HLA-mismatched melanoma or gp100(-) nonmelanoma tumor lines. Importantly, competitive inhibition of lysis of an otherwise susceptible melanoma cell line by cold targets pulsed with known gp100 CD8 T cell epitopes as well as a dose-dependent proliferative response to Th epitopes demonstrates that DCs can process targeted Ag for activation of cytotoxic as well as helper arms of the immune response. Thus, the specific targeting of soluble exogenous tumor Ag to the DC mannose receptor directly contributes to the generation of multiple HLA-restricted Ag-specific T cell responses.

Polysorbate 80 is one of the key components of protein formulations. It primarily inhibits interfacial damage of the protein molecule due to mechanical stress during shipping and handling. However, polysorbate 80 also affects the formulation photostability. Exposure to light of polysorbate 80 aqueous solution results in peroxide generation, which in turn may result in oxidation of the susceptible amino acid residues in the protein molecule. The purpose of this study was to determine if the photostability of our proprietary IgG(1) monoclonal antibody formulation containing polysorbate 80 is affected by the quality (grade/vendor) of polysorbate 80. Following four types of polysorbate 80 were tested: (1) Polysorbate 80 Super-Refined, Mallinckrodt Baker, (2) Polysorbate 80 NF, Mallinckrodt Baker, (3) Polysorbate 80 NF, EMD Chemicals, and (4) Ultra-pure Polysorbate 80 (HX), NOF Corporation. The samples were exposed to light as per ICH guidelines Q1B. The results of the study show that photostability of the antibody formulation is indeed affected by the quality of polysorbate 80. This study underscores the importance of carefully choosing the quality of polysorbate 80 to ensure the robustness of formulation.

A bstract The $ \mathrm{t}\overline{\mathrm{t}} $ production cross section $ \left( {{\sigma_{{\mathrm{t}\overline{\mathrm{t}}}}}} \right) $ is measured in proton-proton collisions at $ \sqrt{s}=7 $ TeV in data collected by the CMS experiment, corresponding to an integrated luminosity of 2.3 fb −1 . The measurement is performed in events with two leptons (electrons or muons) in the final state, at least two jets identified as jets originating from b quarks, and the presence of an imbalance in transverse momentum. The measured value of $ {\sigma_{{\mathrm{t}\overline{\mathrm{t}}}}} $ for a top-quark mass of 172.5 GeV is $ 161.9\pm 2.5\left( {\mathrm{stat}.} \right)_{-5.0}^{+5.1}\left( {\mathrm{syst}.} \right)\pm 3.6\,\left( {\mathrm{lumi}.} \right)\,\mathrm{pb} $ , consistent with the prediction of the standard model.

Polysorbate 80 is widely used in protein formulations to protect protein against agitation-induced aggregation. In this study, we address concerns about residual peroxide present in Polysorbate 80 on protein stability. Residual peroxide may oxidize active pharmaceutical ingredients leading to reduced stability and may ultimately lead to lower potency and efficacy. The effect of Polysorbate 80 concentration on thermal and photostability of monoclonal antibody of the IgG1 subclass (MAb1) was evaluated at Polysorbate 80 concentrations ranging from 0.00% to 1.00% (w/v). MAb1 samples at 5 mg/mL with various Polysorbate 80 concentrations were subjected to accelerated thermal stress by incubation at 25°C, 40°C, and 50°C for a period of 4 weeks and light stress per ICH guideline Q1B, option 1. Our results show that Polysorbate 80 concentration of 1.00% (w/v) adversely affected thermal and photostability of MAb1. This study demonstrates the importance of carefully choosing Polysorbate 80 concentration in protein formulations to prevent destabilizing effect of Polysorbate 80 on thermal and photostability.

ABSTRACT The neutralizing antibody response to the protective antigen (PA) component of anthrax toxin elicited by approved anthrax vaccines is an accepted correlate for vaccine-mediated protection against anthrax. We reasoned that a human anti-PA monoclonal antibody (MAb) selected on the basis of superior toxin neutralization activity might provide potent protection against anthrax. The fully human MAb (also referred to as MDX-1303 or Valortim) was chosen from a large panel of anti-PA human MAbs generated using transgenic mice immunized with recombinant PA solely on the basis of in vitro anthrax toxin neutralization. This MAb was effective in prophylactic and postsymptomatic treatment of rabbits exposed to aerosolized anthrax spores, and a single intramuscular injection of 1 mg/kg of body weight fully protected cynomolgus monkeys challenged with aerosolized anthrax spores. Importantly, MAb 1303 defines a novel neutralizing epitope that requires Fc receptor engagement for maximal activity. F(ab′)2 fragments of MAb 1303, which retain equivalent affinity for PA, are 10- to 100-fold less potent in neutralizing anthrax toxin in vitro. Addition of Fc receptor-blocking antibodies also greatly reduced the activity of MAb 1303. Moreover, we found that the neutralizing activity of mouse, rabbit, and human antisera elicited by PA vaccines was effectively abrogated by blocking Fc receptors. Selection of an anti-PA MAb by using a functional assay that is a surrogate for protection has resulted in the identification of a fully human MAb with potent activity in vivo and uncovered a previously unrecognized mechanism of antibody-mediated toxin neutralization that is important for currently used anthrax vaccines.

A measurement of the t t-bar production cross section in pp collisions at sqrt(s) = 7 TeV is presented. The results are based on data corresponding to an integrated luminosity of 2.3 inverse femtobarns collected by the CMS detector at the LHC. Selected events are required to have one isolated, high transverse momentum electron or muon, large missing transverse energy, and hadronic jets, at least one of which must be consistent with having originated from a b quark. The measured cross section is 158.1 +/- 2.1 (stat.) +/- 10.2 (syst.) +/- 3.5 (lum.) pb, in agreement with standard model predictions.

Beam test results of the radiation tolerance study of chemical vapour deposition (CVD) diamond against different particle species and energies is presented. We also present beam test results on the independence of signal size on incident particle rate in charged particle detectors based on un-irradiated and irradiated poly-crystalline CVD diamond over a range of particle fluxes from 2 kHz/cm2 to 10 MHz/cm2. The pulse height of the sensors was measured with readout electronics with a peaking time of 6 ns. In addition functionality of poly-crystalline CVD diamond 3D devices was demonstrated in beam tests and 3D diamond detectors are shown to be a promising technology for applications in future high luminosity experiments.

A bstract The Compact Linear Collider (CLIC) is a proposed future high-luminosity linear electron-positron collider operating at three energy stages, with nominal centre-of-mass energies $$ \sqrt{s} $$ <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML"> <mml:msqrt> <mml:mi>s</mml:mi> </mml:msqrt> </mml:math> = 380 GeV, 1 . 5 TeV, and 3 TeV. Its aim is to explore the energy frontier, providing sensitivity to physics beyond the Standard Model (BSM) and precision measurements of Standard Model processes with an emphasis on Higgs boson and top-quark physics. The opportunities for top-quark physics at CLIC are discussed in this paper. The initial stage of operation focuses on top-quark pair production measurements, as well as the search for rare flavour-changing neutral current (FCNC) top-quark decays. It also includes a top-quark pair production threshold scan around 350 GeV which provides a precise measurement of the top-quark mass in a well-defined theoretical framework. At the higher-energy stages, studies are made of top-quark pairs produced in association with other particles. A study of t ̄ tH production including the extraction of the top Yukawa coupling is presented as well as a study of vector boson fusion (VBF) production, which gives direct access to high-energy electroweak interactions. Operation above 1 TeV leads to more highly collimated jet environments where dedicated methods are used to analyse the jet constituents. These techniques enable studies of the top-quark pair production, and hence the sensitivity to BSM physics, to be extended to higher energies. This paper also includes phenomenological interpretations that may be performed using the results from the extensive top-quark physics programme at CLIC.

We report on a search for elementary particles with charges much smaller than the electron charge using a data sample of proton-proton collisions provided by the CERN Large Hadron Collider in 2018, corresponding to an integrated luminosity of 37.5 fb$^{-1}$ at a center-of-mass energy of 13 TeV. A prototype scintillator-based detector is deployed to conduct the first search at a hadron collider sensitive to particles with charges ${\leq}0.1e$. The existence of new particles with masses between 20 and 4700 MeV is excluded at 95% confidence level for charges between $0.006e$ and $0.3e$, depending on their mass. New sensitivity is achieved for masses larger than $700$ MeV.

We report on the expected sensitivity of dedicated scintillator-based detectors at the LHC for elementary particles with charges much smaller than the electron charge. The dataset provided by a prototype scintillator-based detector is used to characterize the performance of the detector and provide an accurate background projection. Detector designs, including a novel slab detector configuration, are considered for the data taking period of the LHC to start in 2022 (Run 3) and for the high luminosity LHC. With the Run 3 dataset, the existence of new particles with masses between 10 MeV and 45 GeV could be excluded at 95% confidence level for charges between 0.003 e and 0.3 e, depending on their mass. With the high luminosity LHC dataset, the expected limits would reach between 10 MeV and 80 GeV for charges between 0.0018 e and 0.3 e, depending on their mass.Received 14 April 2021Accepted 12 July 2021DOI:https://doi.org/10.1103/PhysRevD.104.032002Published by the American Physical Society under the terms of the Creative Commons Attribution 4.0 International license. Further distribution of this work must maintain attribution to the author(s) and the published article's title, journal citation, and DOI. Funded by SCOAP3.Published by the American Physical SocietyPhysics Subject Headings (PhySH)Research AreasHypothetical particle physics modelsParticle dark matterTechniquesScintillatorsGeneral PhysicsParticles & Fields

The United States stopped vaccinating against smallpox in 1972 because the risks were judged to outweigh the benefits. The possibility of a terrorist attack using smallpox has led to renewed interest in a vaccination program. Smallpox vaccination carries considerable risks, which may be of greater concern today than in the late 1960s because of the increased prevalence of immunosuppression and atopy in the population. This paper reviews the clinical presentations of major adverse events after vaccination and the rates of occurrence of these events observed in the 1960s. The normal dynamics of the spread of smallpox is slow, and usually only persons who have had close personal contact with an overtly ill patient are affected. There are several preattack vaccination policy options, but immunization of medical workers, especially those who might have close contact with infected patients, is sufficient in the absence of a known threat of a bioterrorist attack or the identification of a smallpox-infected person.

A measurement of the triple-differential cross section (sigma as a function of the photon pt and eta and the jet eta) in photon + jets final states using a data sample from proton-proton collisions at sqrt(s) = 7 TeV is presented. This sample corresponds to an integrated luminosity of 2.14 inverse femtobarns collected by the CMS detector at the LHC. Photons and jets are reconstructed within a pseudorapidity range of abs(eta) < 2.5, and are required to have transverse momenta in the range 40 < pt(gamma) < 300 GeV and pt(jet) > 30 GeV, respectively. The measurements are compared to theoretical predictions from the SHERPA leading-order QCD Monte Carlo event generator and the next-to-leading-order perturbative QCD calculation from JETPHOX. The predictions are found to be consistent with the data over most of the examined kinematic region.

CD27, a member of the TNFR superfamily, is constitutively expressed in most T cells and plays crucial roles in T cell effector functions. The costimulation and antitumor activity of CD27 agonistic Abs have been well documented in mouse models. Clinical testing of a human IgG1 anti-CD27 Ab, varlilumab (clone 1F5), is ongoing in cancer patients. In this study, we set out to further understand CD27 as an immunomodulatory target and to address the mechanism of antitumor efficacy using different IgG isotypes of 1F5 in human CD27-transgenic mice. 1F5mIgG1, the only isotype engaging inhibitory FcγRIIB expressed in B cells, elicited the most potent and broad immune response, but terminal differentiation, exhaustion, and apoptosis in the activated effector T cells were inevitable. Accordingly, this isotype was the most effective in eradicating BCL1 lymphoma but had limited efficacy in s.c. tumors. Conversely, 1F5mIgG2a, which interacts with cells expressing activating FcγRs, led to moderate immune activation, as well as to prominent reduction in the number and suppressive activity of regulatory T cells. These combined mechanisms imparted potent antitumor activity to 1F5mIgG2a, particularly against the s.c. tumors. 1F5hIgG1, varlilumab, showed balanced agonistic activity that was prominent at lower doses and depleting activity that was greater at higher doses. 1F5hIgG1 had good antitumor activity in all tumor models tested. Thus, both agonist and depleting properties contribute to the antitumor efficacy of CD27-targeted immunotherapy, and modulation of these activities in patients may be achieved by varying the dose and regimen.

The AIDA-2020 Trigger Logic Unit (TLU) has been designed to be a flexible and easily configurable unit to provide trigger and control signals to devices employed during test beams, integrating them with the beam telescope. The most recent iteration of the TLU (v1E) has been re-designed within the AIDA-2020 project to integrate with hardware used in beam facilities. Configuration and communication with the TLU are performed over Ethernet. It can be employed as a stand-alone unit or be deployed as part of the EUDAQ2 data acquisition framework, which allows it to connect to a wide range of LHC readout systems. The TLU can operate with a sustained particle rate of 1 MHz and with instantaneous rates up to 20 MHz. In the current firmware iteration, the unit can time-stamp incoming signals with a resolution of 1.5 ns. The hardware, firmware and software designs of the TLU are freely accessible and benefit from constant inputs and upgrades from experienced users. TLU units have already been deployed successfully in beam lines at CERN and DESY.

Abstract Purpose: The oncofetal antigen, human chorionic gonadotropin β subunit (hCGβ), is expressed by a number of carcinomas and is a prognostic indicator in renal, colorectal, bladder, and pancreatic cancers. We describe the development of a novel antibody-based dendritic cell (DC)-targeted cancer vaccine capable of eliciting cellular immune responses directed against hCGβ. Experimental Design: The tumor-associated antigen hCGβ was coupled genetically to a human anti-DC antibody (B11). The resulting fusion protein (B11-hCGβ) was evaluated for its ability to promote tumor antigen-specific cellular immune responses in a human in vitro model. Monocyte-derived human DCs from normal donors were exposed to purified B11-hCGβ, activated with CD40 ligand, mixed with autologous lymphocytes, and tested for their ability to promote hCGβ-specific proliferative and cytotoxic T-lymphocyte responses. Results: B11-hCGβ was found to be a soluble, well-defined, and readily purified product that specifically recognized the human mannose receptor via the B11 antibody portion of the fusion protein. B11-hCGβ functionally promoted the uptake and processing of tumor antigen by DCs, which led to the generation of tumor-specific HLA class I and class II-restricted T-cell responses, including CTLs capable of killing human cancer cell lines expressing hCGβ. Conclusions: Although other hCG vaccines have been shown to be capable of eliciting antibody responses to hCGβ, this is the first time that cellular immune responses to hCGβ have been induced by a vaccine in a human system. This DC-targeted hCGβ vaccine holds promise for the management of a number of cancers and merits additional clinical development.

In the past year, the Linear Collider Flavour Identification (LCFI) Collaboration has taken significant steps towards having a sensor suitable for use in the silicon vertex detector of the International Linear Collider (ILC). The goal of the collaboration is to develop the sensors, electronic systems and mechanical support structures necessary for the construction of a high performance vertex detector and to investigate the contribution such a vertex detector can make to the physics accessible at the ILC. Particular highlights include the delivery and testing of both a second-generation column parallel CCD (CP-CCD), design of the next-generation readout ASIC (CPR2a) and a dedicated ASIC for driving the CP-CCD. This paper briefly describes these and other highlights.

This study demonstrates that arginine is a highly effective solvent additive which significantly reduces the light induced aggregation of four IgG1 type monoclonal antibodies (named as IMC-1A, IMC-1B, IMC-1C and IMC-1D) as measured by size exclusion chromatography. All experiments were performed in a phosphate buffer system containing either sodium chloride or arginine hydrochloride. The protein samples were exposed to light in a photo chamber according to ICH (International Conference on Harmonization) guidelines. Thermal unfolding transition temperature (Tm) of IMC- 1A as determined by differential scanning calorimetry (DSC) was significantly decreased (∼ 3.3°C) in the presence of arginine hydrochloride as compared to in sodium chloride. However, a noticeable increase in thermal stability was observed for IMC-1B, IMC-1C and IMC-1D in the presence of arginine hydrochloride. The photostability of all these molecules was significantly enhanced by arginine hydrochloride and both a direct and inverse correlation was observed between conformational stability and photostability. To our knowledge, this paper for the first time, demonstrates that arginine hydrochloride considerably reduces the light induced aggregation of monoclonal antibodies. Arginine hydrochloride is also known to increase protein solubility and its ability to extensively reduce light induced aggregation makes it a potential solvent additive for the formulation development of therapeutic proteins. Keywords: Photostability, thermal stability, differential scanning calorimetry, aggregation, IgG1

We reviewed the literature on adverse events reported to occur after smallpox vaccination. Nearly one-half of the United States population is vaccinia-naı̈ve and may be at risk for development of serious adverse events. We describe the clinical features of postvaccinial central nervous system disease, progressive vaccinia, eczema vaccinatum, accidental implantations, “generalized vaccinia,” and the common erythematous and/or urticarial rashes. In the 1960s, death occurred approximately once in every million primary vaccinations, with fatalities resulting from progressive vaccinia, postvaccinial encephalitis, and eczema vaccinatum. Death in revaccinees occurred less commonly and almost entirely from progressive vaccinia. In today’s population, death rates might be higher because of the increased prevalence of immune deficiency and atopic dermatitis.

With the commissioning of the LHC in 2010 and upgrades expected in 2015, ATLAS and CMS are planning to upgrade their innermost tracking layers with radiation hard technologies. Chemical Vapor Deposition diamond has been used extensively in beam conditions monitors as the innermost detectors in the highest radiation areas of BaBar, Belle, CDF and all LHC experiments. This material is now being considered as a sensor material for use very close to the interaction region where the most extreme radiation conditions exist. Recently the RD42 collaboration constructed, irradiated and tested polycrystalline and single-crystal chemical vapor deposition diamond sensors to the highest fluences expected at the super-LHC. We present beam test results of chemical vapor deposition diamond up to fluences of 1.8×1016 protons/cm2 illustrating that both polycrystalline and single-crystal chemical vapor deposition diamonds follow a single damage curve. We also present beam test results of irradiated complete diamond pixel modules.

Abstract CD27 is a costimulatory molecule that provides a complementary target to the PD-1/PD-L1 checkpoint axis on T cells. Combining a CD27 agonist antibody with PD-1/PD-L1 blockade has shown synergistic antitumor activity in preclinical models, which led to clinical studies of the combination in cancer patients. We theorized that coupling CD27 costimulation with PD-1/PD-L1 blockade in a bispecific antibody (BsAb) may provide greater immune activating properties than combining the individual mAbs due to enhanced CD27 activation by cross-linking through PD-L1 and Fc receptors. To test this approach, we developed CDX-527, a tetravalent PD-L1xCD27 IgG1-scFv BsAb. CDX-527 potently inhibits PD-1 signaling and induces CD27-mediated T cell costimulation through PD-L1 cross-linking. In mixed lymphocyte reaction assays, CDX-527 is more potent than the combination of the parental antibodies, suggesting that cross-linking through both Fc receptors and PD-L1 results in enhanced CD27 agonist activity. CDX-527 was shown to mediate effector function against tumor cells overexpressing either CD27 or PD-L1. In human CD27 transgenic mice, we observed that antigen-specific T cell responses to a vaccine are greatly enhanced with a surrogate PD-L1xCD27 BsAb. Furthermore, the BsAb exhibits greater antitumor activity than the combination of the parental antibodies in a syngeneic lymphoma model. A pilot study of CDX-527 in cynomolgus macaques confirmed a mAb-like pharmacokinetic profile without noted toxicities. These studies demonstrate that CDX-527 effectively combines PD-1 blockade and CD27 costimulation into one molecule that is more potent than combination of the parental antibodies providing the rationale to advance this BsAb toward clinical studies in cancer patients.

Vaccine therapy is attractive for prostate cancer patients because the tumor is slow growing (allowing time to augment host responses) and occurs in an older population less likely to tolerate more toxic treatments. We have constructed an expression vector based on a monoclonal antibody (mAb) that targets the high affinity receptor for IgG (FcγRI, CD64) which is exclusively expressed on myeloid cells including dendritic cells (DC). The heavy chain of mAb H22 CH2 and CH3 domains were removed and replaced with the gene for prostate specific antigen (PSA). Using that vector, we have constructed and purified FPH22.PSA, a fusion protein that targets PSA to FcγRI on antigen presenting cells (APC). This fusion protein has an apparent molecular mass of 80–83 kDa, binds to FcγRI with high affinity and expresses PSA. We demonstrate that FPH22.PSA targeted PSA was internalized and processed by the human myeloid THP-1 cell line resulting in presentation of MHC class I-associated PSA peptides and lysis of THP-1 by PSA-specific human CTL. Moreover, pretreatment of THP-1 cells with antibodies to block either FcγRI or MHC class I, blocked lysis indicating that targeting to FcγRI results in presentation of exogenous antigen on MHC class I molecules. These data demonstrate that FPH22.PSA was processed in such a manner by the myeloid cell line to allow for presentation of immunodominant peptides in MHC class I molecules and suggests that uptake of antigen via FcγRI results in cross-priming.

I- CreI is a member of the LAGLI-DADG family of homing nucleases; however, unlike most members of this family it contains only a single copy of this signature motif. I- CreI was over-expressed in Escherichia coli, and a simple purification protocol developed that gave reasonably pure protein in high yield. Size-exclusion chromatography and chemical cross-linking indicated that the protein is a dimer in solution. DNA cleavage by I- CreI was absolutely dependent on Mg2+(or Mn2+), and was inhibited by monovalent cations. I- CreI displayed a surprisingly high temperature optimum (>50 degrees C), with full activity occurring even at 70 degrees C. Interestingly, SDS was needed for efficient release of the cleavage products from the protein, indicating formation of very stable DNA-protein complexes. In contrast to these robust characteristics, purified I- CreI was unstable; however, it could be stabilized by the addition of either target or non-target DNA. Mobility shift assays revealed that I- CreI binds to DNA in the absence of Mg2+. Hydroxyl radical footprinting showed that I- CreI strongly protected the backbone of a continuous stretch of at least 12 nt on each strand that were shifted, relative to each other, by 2 bp in the 3'direction. Methylation protection and interference analyses were also performed, and together with the hydroxyl radical footprinting, indicate that I- CreI binds in both the major and minor grooves of its target DNA.

The production of a standard model Higgs boson in association with a top quark pair at the upcoming high luminosity run ( 15 fb(-1) integrated luminosity) of the Fermilab Tevatron ( square root of s = 2.0 TeV) is revisited. For Higgs masses below 140 GeV we demonstrate that the production cross section times branching ratio for H-->bb macro decays yields a significant number of events and that this mode is competitive with and complementary to the searches using pp(macro) -->WH,ZH associated production. For higher mass Higgs bosons the H-->W(+)W(-) decays are more difficult but have the potential to provide a few spectacular events.

The precision measurements envisaged at the International Linear Collider (ILC) depend on excellent instrumentation and reconstruction software. The correct identification of heavy flavour jets, placing unprecedented requirements on the quality of the vertex detector, will be central for the ILC programme. This paper describes the LCFIVertex software, which provides tools for vertex finding and for identification of the flavour and charge of the leading hadron in heavy flavour jets. These tools are essential for the ongoing optimisation of the vertex detector design for linear colliders such as the ILC. The paper describes the algorithms implemented in the LCFIVertex package as well as the scope of the code and its performance for a typical vertex detector design.

The hydrophobic region of the signal peptide of the OmpA protein of the Escherichia coli outer membrane was extensively altered in its hydrophobicity and predicted secondary structure by site-specific mutagenesis. The mutated signal peptides were fused to nuclease A from Staphylococcus aureus, and the function of the signal peptide was examined by measuring the rate of processing of the signal peptide. Six of the 12 mutated signal peptides in the nuclease hybrid were processed faster than the wild-type. In particular, the processing of the mutated signal peptide in which the alanine residue at position 9 was substituted with a valine residue was enhanced almost twofold over the processing of the wild-type signal peptide. In addition, the production of nuclease A fused with this mutated signal peptide also increased twofold. However, these effects were not observed when the mutated signal peptide was fused to TEM beta-lactamase. Analysis of the present mutations suggests that both overall hydrophobicity and distinct structural requirements in the hydrophobic region have important roles in signal peptide function.

Low levels of alanine to serine sequence variants were identified in an IgG4 monoclonal antibody by ultra/high performance liquid chromatography and tandem mass spectrometry. The levels of the identified sequence variants A183S and A152S, both in the light chain, have been determined to be 7.8–9.9% and 0.5–0.6%, by extracted ion currents of the tryptic peptides L16 and L14, respectively. The A183S variant was confirmed through tryptic map spiking experiments using synthetic peptide, SDYEK, which incorporated Ser at the position of native Ala in the tryptic peptide L16. Both mutations were also observed by endoproteinase Asp-N peptide mapping. The variant level of A183S was also quantified by LC–UV with detection at 280 nm and fluorescence detection of tyrosine residues on the tryptic peptides. The results from LC–MS, UV, and fluorescence detection are in close agreement with each other. The levels of the sequence variants are comparable among the antibody samples manufactured at different scales as well as locations, indicating that the variants’ levels are not affected by manufacture scale or locations. DNA sequencing of the master cell bank revealed the presence of mixed bases at position 183 encoding both wild and mutated populations, whereas bases encoding the minor sequence variant at position 152 were not detected. The root cause for A152S mutation is not yet clearly understood at this moment.

Abstract The PLUME (Pixelated Ladder with Ultra-Low Material Embedding) Collaboration is developing ultra-light ladders for the vertex detector for a future linear collider. The double-sided ladder will integrate the sensors, readout infrastructure and mechanical supports with the aim of total material budget of 0.3% of radiation length. The requirement of as light as possible construction is driven by physics, in particular by measurements requiring determination of the quark charge sign. The first prototype ladders were prepared and tested in the beam. The alignment issues for the ladders will be tested within the AIDA (Advanced European Infrastructures for Detectors at Accelerators) EU FP7 project.

The ratio of the production cross sections times branching fractions $$ \left(\sigma \left({\mathrm{B}}_{\mathrm{c}}^{\pm}\right)\mathrm{\mathcal{B}}\left({\mathrm{B}}_{\mathrm{c}}^{\pm}\to \mathrm{J}/\psi {\pi}^{\pm}\right)\right)/\left(\sigma \left({\mathrm{B}}^{\pm}\right)\mathrm{\mathcal{B}}\left({\mathrm{B}}^{\pm}\to \mathrm{J}/\psi {K}^{\pm}\right)\right) $$ is studied in proton-proton collisions at a center-of-mass energy of 7 TeV with the CMS detector at the LHC. The kinematic region investigated requires B c ± and B± mesons with transverse momentum p T > 15 GeV and rapidity |y| < 1.6. The data sample corresponds to an integrated luminosity of 5.1 fb−1. The ratio is determined to be $$ \left[0.48\pm 0.05\left(\mathrm{stat}\right)\pm 0.03\left(\mathrm{syst}\right)\pm 0.05\ \left({\tau}_{{\mathrm{B}}_{\mathrm{c}}}\right)\right]\% $$ . The B c ± → J/ψπ ± π ± π ∓ decay is also observed in the same data sample. Using a model-independent method developed to measure the efficiency given the presence of resonant behaviour in the three-pion system, the ratio of the branching fractions $$ \mathrm{\mathcal{B}}\left({\mathrm{B}}_{\mathrm{c}}^{\pm}\to \mathrm{J}/\psi {\pi}^{\pm }{\pi}^{\pm }{\pi}^{\mp}\right)/\mathrm{\mathcal{B}}\left({\mathrm{B}}_{\mathrm{c}}^{\pm}\to \mathrm{J}/\psi {\pi}^{\pm}\right) $$ is measured to be $$ 2.55\pm 0.80\left(\mathrm{stat}\right)\pm 0.33{\left(\mathrm{syst}\right)}_{-0.01}^{+0.04}\left({\tau}_{B_c}\right) $$ , consistent with the previous LHCb result.

Pace et al. (1995) [1] recommended an equation used to predict extinction coefficient of a protein. However, no antibody data was included in the development of this equation. The main objective of this study was to therefore investigate how the predicted value of the extinction coefficient is comparable to the experimentally determined extinction coefficient of antibodies measured by the Edelhoch method. We have measured the extinction coefficients (ɛ) of 13 IgG1 monoclonal antibodies (mAbs) in phosphate buffer at pH 7.2. The maximum variability in the experimentally measured extinction coefficient of a given mAb molecule was found to be about 2%. Experimentally determined extinction coefficients of all mAbs were found to be lower than the predicted value, with the maximum difference found to being 4.7%. The highest and lowest values of experimental extinction coefficient among the thirteen IgG1 monoclonal antibodies obtained were 230525.9 M−1 cm−1 (i.e. 1.55 (mg/ml)−1 cm−1) and 191,411.6 M−1 cm−1 (i.e. 1.29 (mg/ml)−1 cm−1). A difference of <3% (with respect to mean value) was observed between the experimental and predicted values of the extinction coefficient. A comprehensive analysis and interpretation of the comparison of the predicted and experimentally determined extinction coefficient by the Edelhoch method is discussed in terms of structural characterization and accessible surface area (ASA).

The cross section for the production of Zγ in proton-proton collisions at 8 TeV is measured based on data collected by the CMS experiment at the LHC corresponding to an integrated luminosity of 19.5 fb−1. Events with an oppositely-charged pair of muons or electrons together with an isolated photon are selected. The differential cross section as a function of the photon transverse momentum is measured inclusively and exclusively, where the exclusive selection applies a veto on central jets. The observed cross sections are compatible with the expectations of next-to-next-to-leading-order quantum chromodynamics. Limits on anomalous triple gauge couplings of ZZγ and Zγγ are set that improve on previous experimental results obtained with the charged lepton decay modes of the Z boson.

Abstract We have measured the radiation tolerance of poly-crystalline and single-crystalline diamonds grown by the chemical vapor deposition (CVD) process by measuring the charge collected before and after irradiation in a 50 m pitch strip detector fabricated on each diamond sample. We irradiated one group of sensors with 800 MeV protons, and a second group of sensors with 24 GeV protons, in steps, to protons cm −2 and protons cm −2 respectively. We observe the sum of mean drift paths for electrons and holes for both poly-crystalline CVD diamond and single-crystalline CVD diamond decreases with irradiation fluence from its initial value according to a simple damage curve characterized by a damage constant for each irradiation energy and the irradiation fluence. We find for each irradiation energy the damage constant, for poly-crystalline CVD diamond to be the same within statistical errors as the damage constant for single-crystalline CVD diamond. We find the damage constant for diamond irradiated with 24 GeV protons to be and the damage constant for diamond irradiated with 800 MeV protons to be . Moreover, we observe the pulse height decreases with fluence for poly-crystalline CVD material and within statistical errors does not change with fluence for single-crystalline CVD material for both 24 GeV proton irradiation and 800 MeV proton irradiation. Finally, we have measured the uniformity of each sample as a function of fluence and observed that for poly-crystalline CVD diamond the samples become more uniform with fluence while for single-crystalline CVD diamond the uniformity does not change with fluence.

Oligonucleotide-directed site-specific mutagenesis was used to study the structure-function relationship of the positively charged amino terminus of the Escherichia coli outer membrane protein OmpA signal peptide. Mutations were isolated which reduced the overall charge of the amino-terminal region from +2 (wild type) to +1, 0, and -1, as well as one mutation from Thr to Ser at position 4. DNA encoding the wild type and mutant OmpA signal peptides was then fused in-frame to DNA encoding the mature regions of Staphylococcus aureus nuclease A and TEM beta-lactamase. In the case of both the beta-lactamase and nuclease fusions, normal processing was no longer observed when the charge at the amino terminus was reduced to zero or made negative. Differences between the two hybrid proteins were observed in the case of the Thr to Ser mutation. As expected, this mutation had no effect on the beta-lactamase hybrid; however, the processing rate of the nuclease hybrid protein was reduced to nearly one-half. Furthermore, this effect was essentially reversed when a Lys residue at position 3 was deleted. A model is presented which explains the differing effects of a signal peptide mutation on the secretion of different hybrid proteins based on kinetic differences in the translocation of the nuclease and beta-lactamase proteins.

We measured the radiation tolerance of commercially available diamonds grown by the Chemical Vapor Deposition process by measuring the charge created by a 120 GeV hadron beam in a 50 μm pitch strip detector fabricated on each diamond sample before and after irradiation. We irradiated one group of samples with 70 MeV protons, a second group of samples with fast reactor neutrons (defined as energy greater than 0.1 MeV), and a third group of samples with 200 MeV pions, in steps, to (8.8±0.9) × 1015 protons/cm2, (1.43±0.14) × 1016 neutrons/cm2, and (6.5±1.4) × 1014 pions/cm2, respectively. By observing the charge induced due to the separation of electron-hole pairs created by the passage of the hadron beam through each sample, on an event-by-event basis, as a function of irradiation fluence, we conclude all datasets can be described by a first-order damage equation and independently calculate the damage constant for 70 MeV protons, fast reactor neutrons, and 200 MeV pions. We find the damage constant for diamond irradiated with 70 MeV protons to be 1.62±0.07(stat)±0.16(syst)× 10-18 cm2/(p μm), the damage constant for diamond irradiated with fast reactor neutrons to be 2.65±0.13(stat)±0.18(syst)× 10-18 cm2/(n μm), and the damage constant for diamond irradiated with 200 MeV pions to be 2.0±0.2(stat)±0.5(syst)× 10-18 cm2/(π μm). The damage constants from this measurement were analyzed together with our previously published 24 GeV proton irradiation and 800 MeV proton irradiation damage constant data to derive the first comprehensive set of relative damage constants for Chemical Vapor Deposition diamond. We find 70 MeV protons are 2.60 ± 0.29 times more damaging than 24 GeV protons, fast reactor neutrons are 4.3 ± 0.4 times more damaging than 24 GeV protons, and 200 MeV pions are 3.2 ± 0.8 more damaging than 24 GeV protons. We also observe the measured data can be described by a universal damage curve for all proton, neutron, and pion irradiations we performed of Chemical Vapor Deposition diamond. Finally, we confirm the spatial uniformity of the collected charge increases with fluence for polycrystalline Chemical Vapor Deposition diamond, and this effect can also be described by a universal curve.

Unrecoverable internal failures of modules in the CDF Run2 Silicon detector have been observed since its installation in early 2001. A fraction of these failures has been categorized as infant mortality. Other failures occurring later were strongly correlated with fixed trigger conditions. These failures are explained by wire-bonds breaking due to fatigue stress induced by resonant vibration. These resonant vibrations are a direct consequence of the oscillating Lorentz forces induced by the 1.4T magnetic field on wire-bonds carrying non-DC current. Changes have been implemented in data-taking procedures in order to minimize the occurrences of such failures and to prolong the lifetime of the detector itself. A more general analysis of the topic has been pursued. Changes in the packaging and assembly processes for future applications have been investigated.

Diamond devices have now become ubiquitous in the LHC experiments, finding applications in beam background monitoring and luminosity measuring systems. This sensor material is now maturing to the point that the large pads in existing diamond detectors are being replaced by highly granular tracking devices, in both pixel and strip configurations, for detector systems that will be used in Run II at the LHC and beyond. The RD42 collaboration has continued to seek out additional diamond manufacturers and quantify the limits of the radiation tolerance of this material. The ATLAS experiment has recently installed, and is now commissioning a fully-fledged pixel tracking detector system based on diamond sensors. Finally, RD42 has recently demonstrated the viability of 3D biased diamond sensors that can be operated at very low voltages with full charge collection. These proceedings describe all of these advances.

On the basis of the biophysical studies on the synthetic mutant (Ile-8 4 Asn) OmpA signal peptide in the preceding paper (Hoyt,

In this LOI we propose a dedicated experiment that would detect milli-charged particles produced by pp collisions at LHC Point 5. The experiment would be installed during LS2 in the vestigial drainage gallery above UXC and would not interfere with CMS operations. With 300 fb$^{-1}$ of integrated luminosity, sensitivity to a particle with charge $\mathcal{O}(10^{-3})~e$ can be achieved for masses of $\mathcal{O}(1)$ GeV, and charge $\mathcal{O}(10^{-2})~e$ for masses of $\mathcal{O}(10)$ GeV, greatly extending the parameter space explored for particles with small charge and masses above 100 MeV.

Detectors based on Chemical Vapor Deposition (CVD) diamond have been used extensively and successfully in beam conditions/beam loss monitors as the innermost detectors in the highest radiation areas of Large Hadron Collider (LHC) experiments. The startup of the LHC in 2015 brought a new milestone where the first polycrystalline CVD (pCVD) diamond pixel modules were installed in an LHC experiment and successfully began operation. The RD42 collaboration at CERN is leading the effort to develop polycrystalline CVD diamond as a material for tracking detectors operating in extreme radiation environments. The status of the RD42 project with emphasis on recent beam test results is presented.

Immunoglobulin-like transcript 4 (ILT4) is an immunosuppressive molecule predominantly expressed on myeloid cells. Recent studies combining ILT4 suppression with programmed cell death 1 (PD-1)/programmed cell death ligand 1 (PD-L1) blockade have shown promising signs of activity in immune checkpoint inhibitor refractory patients. We theorized that coupling ILT4 and PD-1/PD-L1 blockade in a bispecific antibody (bsAb) may provide greater immune activating properties than combining the individual mAbs due to enhanced bridging of APCs to T cells. To test this approach, we developed CDX-585, a tetravalent ILT4xPD-1 IgG1-scFv bsAb from novel PD-1 and ILT-4 mAbs. CDX-585 is a potent antagonist of both PD-1 and ILT4. CDX-585 promotes M1 macrophage polarization and enhances pro-inflammatory cytokine secretion in response to lipopolysaccharide or CD40 agonist mAb treatment. In mixed lymphocyte reaction (MLR) assays, CDX-585 is more potent than the combination of parental antibodies. In a humanized NCG mouse SK-MEL-5 tumor model, CDX-585 exhibits greater antitumor activity than the combination of parental mAbs. A pilot study of CDX-585 in cynomolgus macaques confirmed a mAb-like pharmacokinetic profile without noted toxicities. These studies demonstrate that CDX-585 effectively combines ILT4 and the PD-1 blockade into one molecule that is more potent than the combination of the parental antibodies, providing the rationale to advance this bsAb into clinical studies.

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTDesign and Evaluation of a Thrombin-Activable Plasminogen ActivatorWen Pin Yang, Joel Goldstein, Roman Procyk, Gary R. Matsueda, and Shyh Yu ShawCite this: Biochemistry 1994, 33, 8, 2306–2312Publication Date (Print):March 1, 1994Publication History Published online1 May 2002Published inissue 1 March 1994https://doi.org/10.1021/bi00174a043Request reuse permissionsArticle Views74Altmetric-Citations16LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit PDF (2 MB) Get e-Alertsclose Get e-Alerts

Expression of the type I receptor for Fc domain of immunoglobulin (Ig)G (Fc gammaRI or CD64) is restricted to myeloid effector cells, such as monocytes, macrophages and a subset of dendritic cells. Previous work has indicated a role for Fc gammaRI in antibody-dependent phagocytosis and lysis of tumour cells. We hypothesised that tagging of tumour cells with an anti-Fc gammaRI single chain Fv (sFv) may facilitate targeting to this receptor on effector cells, thereby initiating tumour cytotoxicity. A vector encoding the sFv for an Fc gammaRI-specific antibody (H22), linked to the transmembrane domain of platelet-derived growth factor was constructed. Transfected tumour cells expressed high surface levels of functional H22-sFv, which greatly enhanced susceptibility for phagocytosis and lysis by monocytes and macrophages. The expression of H22-sFv evoked the ability of tumour cells to directly activate monocytes, as evidenced by phosphorylation of mitogen-activated protein kinase and secretion of the inflammatory cytokines interleukin (IL)-1beta, tumour necrosis factor-alpha and IL-6. Moreover, growth of tumour cells in mice expressing H22-sFv was profoundly delayed (or absent) in transgenic mice expressing human Fc gammaRI. These results demonstrated that tumour cells can be readily modified to activate cell effector mechanisms, a strategy that may be useful for in vivo targeting in patients.

The strategy for a comparability assessment is developed on a hierarchical risk-based approach. Critical analysis of physicochemical and biological characterization assays is essential for the development of a good comparability protocol. Therefore, selection and sensitivity of these assays is very important. This article discusses a case study to evaluate the sensitivity of various methods in a comparability assessment of three lots of an IgG1 monoclonal antibody (mAb). Analysis with eighteen methods demonstrated that only six of the methods were sensitive enough to show a measurable difference of comparability under accelerated conditions (40°C). Samples stored at 4°C were found to be comparable by all methods. A brief comparison of the results of biochemical and functional assays with biophysical analysis is discussed. Basic principles, applications, strength, and limitations of different biophysical methods are also discussed here. Keywords: Comparability, aggregation, degradation, stability, differential scanning calorimetry, fluorescence, light scattering, IgG, monoclonal antibody (mAb), biophysical analysis, physicochemical characterization, in vivo, pharmacokinetic (PK), biophysical techniques, clinical trials.

In sight of the luminosity increase of the High Luminosity-LHC (HL-LHC), most experiments at the CERN Large Hadron Collider (LHC) are planning upgrades for their innermost layers in the next 5–10 years. These upgrades will require more radiation tolerant technologies than exist today. Usage of Chemical Vapor Deposition (CVD) diamond as detector material is one of the potentially interesting technologies for the upgrade. CVD diamond has been used extensively in the beam condition monitors of BaBar, Belle, CDF and all LHC experiments. Measurements of the radiation tolerance of the highest quality polycrystalline CVD material for a range of proton energies, pions and neutrons obtained with this material are presented. In addition, new results on the evolution of various semiconductor parameters as a function of the dose rate are described.

In the present study, results towards the development of a 3D diamond sensor are presented. Conductive channels are produced inside the sensor bulk using a femtosecond laser. This electrode geometry allows full charge collection even for low quality diamond sensors. Results from testbeam show that charge is collected by these electrodes. In order to understand the channel growth parameters, with the goal of producing low resistivity channels, the conductive channels produced with a different laser setup are evaluated by Raman spectroscopy.

Abstract CD40, a member of the tumor necrosis factor receptor (TNFR) superfamily, is essential for activating both innate and adaptive immune systems. CD40 is expressed by antigen-presenting cells (APC) including dendritic cells and B cells. The engagement by its natural ligand (CD40L) on T cells activates APC thus enhancing immune responses. CD40 is also expressed on many cancer cells, in particular B cell lymphomas, and agonist CD40 antibodies can lead to apoptosis and impaired growth of cancer cells due to direct signaling effects. There are several CD40 agonist antibodies in clinical development, which are differentiated by their level of agonist activity, route of administration, and associated side effect profile. We have generated a panel of CD40-specific fully human antibodies derived from human immunoglobulin transgenic mice with the aim to select a clinical candidate with strong agonist activity and a safety profile that allows for systemic dosing. CDX-1140 is a human IgG2 antibody that stimulates CD40 signaling without the requirement for cross-linking or Fc receptor interactions. CDX-1140 is a potent stimulator of human B cells and dendritic cells in vitro, and has potent direct anti-tumor activity against B cell lymphoma cell lines transplanted into immunocompromised mice. To demonstrate the contribution of immune cell activation to the anti-lymphoma activity in vivo, we included human PBMCs in subsequent xenograft studies. Using human B cell lymphoma cell lines Raji and Ramos as tumor models and severe combined immunodeficiency mice (SCID-beige) as hosts, we found that addition of human PBMC alone had a minor impact on tumor growth (median survival 32 days, compared to 26 days with no treatment, p&lt;0.01). Administration of CDX-1140 alone showed partial anti-tumor activity (median survival 39 days, p&lt; 0.001), while the combination of human PBMCs and CDX-1140 provided a significant increase in survival with all mice alive at day 45. These studies provide evidence that CDX-1140 is able to induce in vivo activation and anti-tumor activity of effector cells derived from injected human PBMC, in addition to the direct signaling effects on lymphoma cells. We are investigating anti-lymphoma efficacy of combining CDX-1140 with other immunotherapies including varlilumab, our CD27 agonist antibody, which is currently in clinical development. Together with our prior preclinical studies characterizing this antibody, these results suggest that CDX-1140 is a promising candidate for clinical development in patients with lymphomas. Disclosures He: Celldex Therapeutics, Inc.: Employment. Testa:Celldex Therapeutics, Inc.: Employment. Anna:Celldex Therapeutics, Inc.: Employment. Jeffery:Celldex Therapeutics, Inc.: Employment. Sisson:Celldex Therapeutics, Inc.: Employment. Vitale:Celldex Therapeutics, Inc.: Employment. O'Neill:Celldex Therapeutics, Inc.: Employment. Crocker:Celldex Therapeutics, Inc.: Employment. Widger:Celldex Therapeutics, Inc.: Employment. Goldstein:Celldex Therapeutics, Inc.: Employment. Marsh Jr.:Celldex Therapeutics, Inc.: Employment. Keler:Celldex Therapeutics: Employment, Equity Ownership.

The CBC3 is the latest version of the CMS Binary Chip ASIC for readout of the outer radial region of the upgraded CMS Tracker at HL-LHC.This 254-channel, 130nm CMOS ASIC is designed to be bump-bonded to a substrate to which sensors will be wire-bonded.It will instrument double-layer 2S-modules, consisting of two overlaid silicon microstrip sensors with aligned microstrips.On-chip logic identifies first level trigger primitives from high transversemomentum tracks by selecting correlated hits in the two sensors.Delivered in late 2016, the CBC3 has been under test for several months, including X-ray irradiations and SEU testing.Results and performance are reported.

Prodrug esters of the indolocarbazole CEP-751 (KT-6587) were prepared with the goal of identifying water soluble, stable but cleavable forms for intravenous dosing. A dipeptide proform Lys-β-Ala (16, CEP-2563/KT-8391) was identified for advancement to clinical trials.

We have measured the radiation tolerance of commercially available diamonds grown by the Chemical Vapor Deposition process by measuring the charge created by a 120 GeV hadron beam in a 50 µm pitch strip detector fabricated on each diamond sample before and after irradiation.We irradiated one group of samples with 70 MeV protons, a second group of samples with fast reactor neutrons (defined as energy greater than 0.1 MeV) and a third group of samples with 200 MeV pions, in steps, to (8.8 ± 0.9) × 10 15 protons/cm 2 , (1.43 ± 0.14) × 10 16 neutrons/cm 2 and (6.5 ± 0.5) × 10 14 pions/cm 2 respectively.By observing the charge induced due to the separation of electron-hole pairs created by the passage of the hadron beam through each sample, on an event-by-event basis, as a function of irradiation fluence, we conclude all data sets can be described by a first order damage equation and independently calculate the damage constant for 70 MeV protons, fast reactor neutrons and 200 MeV pions.We find the damage constant for diamond irradiated with 70 MeV protons to be 1.61 ± 0.07 (stat) ± 0.15 (syst) × 10 -18 cm 2 /(p µm), the damage constant for diamond irradiated with fast reactor neutrons to be 2.65 ± 0.13 (stat) ± 0.16 (syst) × 10 -18 cm 2 /(n µm) and the damage constant for diamond irradiated with 200 MeV pions to be 2.0 ± 0.2 (stat) ± 0.5 (syst) × 10 -18 cm 2 /(π µm).The damage constants from this measurement were analyzed together with our previously published 24 GeV proton irradiation and 800 MeV proton irradiation damage constant data to derive the first comprehensive set of relative damage constants for Chemical Vapor Deposition diamond.We find 70 MeV protons are 2.60 ± 0.27 times more damaging than 24 GeV protons, fast reactor neutrons are 4.27 ± 0.34 times more damaging than 24 GeV protons and 200 MeV pions are 3.2 ± 0.8 more damaging than 24 GeV protons.We also observe the measured data can be described by a universal damage curve for all proton, neutron and pion irradiations we have performed of Chemical Vapor Deposition diamond.Finally, we confirm the FWHM/MP ratio of the signal spectrum, a measure of the spatial uniformity of the collected charge, decreases with fluence for polycrystalline Chemical Vapor Deposition diamond and this effect can also be described by a universal curve.

The status of the Silicon Microvertex Detector (SMD) and its installation into the LEP-I.3experiment are presented, highiighting novel features and sophisticated techniques.Preliminary results based on 1993 data are given and compared with Monte Carlo predictions, to understand the detector performances and its tracking capabilities.

The conclusions reached in the case report of congenital vaccinia by Harley and Gillespie are open to question. Eighteen cases of congenital vaccinia have been desiribed, none with congenital malformations. Two infants survived; both mothers were vaccinated at about 32 weeks' gestation. In studies involving a total of 7,886 vaccinated pregnant women and 8,158 unvaccinated pregnant women, there was no evidence of an increased incidence of congenital malformations in the vaccinated subjects. There is a lack of objective evidence that the congenital abnormalities described in this patient were related to his mother's smallpox vaccination.

The CBC3 is the latest version of the CMS Binary Chip for readout of the outer radial region of the upgraded CMS Tracker at the High Luminosity LHC. This 254-channel, 130 nm CMOS ASIC is designed to be bump-bonded to a substrate to which sensors will be wire-bonded. It will instrument double-layer 2S-modules, containing two overlaid silicon microstrip sensors, aligned with a parallel orientation. On-chip logic identifies Level-1 trigger primitives from high transverse-momentum tracks by selecting correlated clusters in the two sensors. The CBC3 was delivered in late 2016; wafer probing and performance tests have been carried out. Several prototype modules using the CBC3 have been produced and tested in the lab and in different beams. The results show that the CBC3 satisfies CMS requirements and only small corrections are needed for the final version of the chip for production.

In this LOI we propose a dedicated experiment that would detect "milli-charged" particles produced by pp collisions at LHC Point 5. The experiment would be installed during LS2 in the vestigial drainage gallery above UXC and would not interfere with CMS operations. With 300 fb$^{-1}$ of integrated luminosity, sensitivity to a particle with charge $\mathcal{O}(10^{-3})~e$ can be achieved for masses of $\mathcal{O}(1)$ GeV, and charge $\mathcal{O}(10^{-2})~e$ for masses of $\mathcal{O}(10)$ GeV, greatly extending the parameter space explored for particles with small charge and masses above 100 MeV.

Many children with recurrent sinopulmonary infections fail to mount an adequate humoral response following immunization with polysaccharide antigens. At present there are no controlled studies comparing responses to pneumococcal immunization in children with recurrent infections and a healthy, age-matched cohort. Immunological evaluation was performed on 66 children with recurrent sinopulmonary infections, aged 2-5 years (mean 3.06 +/- 0.92). A control group included 28 healthy, age-matched controls (mean 3.14 +/- 0.88 years). Both groups were immunized with 23 valent pneumococcal vaccine, and titers were measured before and 4 weeks after immunization. Antibody levels to 12 pneumococcal serotypes were measured via radioimmunoassay. Geometric preimmunization mean titers in the control group were 215.5 +/- 157 ngAbN/ml rising to 989.5 +/- 745 ngAbN/ml compared to 77.71 +/- 38.4 ngAbN/ml increasing to 446.7 +/- 406 ngAbN/ml in the study group (p < .05). Serotypes 3, 4, 7F, 8, 9N, and 18C were the most immunogenic, while serotypes 6A and 14 were the least. Overall, the control group responded to 7.71 +/- 1.24 serotypes versus 5.1 +/- 2.0 in the study group (p < .05), where postimmunization titers at least doubled and rose to > or = 300 ngAbN/ml. All controls responded to at least five or more serotypes, 26/28 responded to 6 or more. In contrast, only 38/66 (57%) of study patients responded to five or more serotypes, and only 27/66 (41%) responded to at least 6 of 12. Preimmunization titers of greater than 300 ngAbN/ml were present in 30% (102/336) of the control serotypes; however, only 53 of these (52%) doubled post immunization; 22% of the elevated titers decreased post immunization. Markedly elevated titers > or = 500 ngAbN/ml were present in 20% (69/336) of the preimmunization serotypes, only 39% of these doubled post immunization. Twenty-three valent pneumococcal vaccine is immunogenic in young, healthy children. A significant percentage of children with recurrent sinopulmonary infections fail to produce adequate serotype specific antibodies following pneumococcal immunization.

The limulus amebocyte lysate test for endotoxin was done on samples of blood from 22 well babies and 33 neonates in an intensive-care nursery. The objective was to determine whether falsely positive test results occurred in samples from newborn infants during acquisition of usual bowel flora. One neonate had a transiently positive limulus amebocyte lysate test; he had clinical signs of sepsis, but no bacteremia could be documented. Unlike the nitroblue tetrazolium test, the limulus amebocyte lysate test does not appear to be regularly positive in neonates. The sensitivity of the test in detection of endotoxemia associated with gram-negative sepsis in the neonate remains to be determined in a large prospective study.

The pain-reducing and calming properties of nitrous oxide mixed with oxygen were evaluated with the use of placebo and standard control groups. Subjects under nitrous oxide had higher pain thresholds and a higher tolerance for shock than placebo and control subjects. They also reported themselves as more relaxed than did placebo and control subjects.

The in situ storage image sensor (ISIS) is a monolithic active pixel sensor with memory cells in each pixel. The memory cells are implemented as a CCD register. This and other features of the sensor make the ISIS an excellent device for detectors that will be used at the International linear collider (ILC), an electron-positron accelerator with a proposed centre-of-mass energy of around 500 GeV. The sensor can be made very thin while retaining a high signal-to-noise ratio. The memory cells can be read out between bunch trains at a relatively low clock speed, hence limiting power consumption. The actual signal charge is stored in potential wells and not on capacitors, minimizing the sensitivity to electromagnetic interference. This paper presents the results of the first beam test of the first ISIS prototype. The measurements made include the signal-to-noise ratio, position resolution and efficiency.

Formulation robustness is critical to product quality. A challenge in designing robustness into biologic formulations is the absence of one global parameter, which allows equitable comparison across formulations and quality measures. The new measure introduced here derived from accelerated testing data allows for the design of globally optimal robust formulations. The measure is easy to understand and also numerically and computationally stable when performing optimizations. The measure termed here as robustness index (RobX) score is a relative measure allowing for global optimization across multiple critical quality attributes. Described here are the calculation methods, a discussion of the measure characteristics, and a brief example of using RobX to find a global optimal formulation across multiple quality characteristics within the framework of a mixture experimental design. The data presented are actual stability data generated during a formulation experimental design for a monoclonal antibody. We anticipate this approach will provide a good general measure of drug stability and will help to accelerate the drug development process. We believe it will improve formulation robustness, long-term stability, and the delivery of quality drug products to the patient. © 2011 Wiley Periodicals, Inc. and the American Pharmacists Association. Formulation robustness is critical to product quality. A challenge in designing robustness into biologic formulations is the absence of one global parameter, which allows equitable comparison across formulations and quality measures. The new measure introduced here derived from accelerated testing data allows for the design of globally optimal robust formulations. The measure is easy to understand and also numerically and computationally stable when performing optimizations. The measure termed here as robustness index (RobX) score is a relative measure allowing for global optimization across multiple critical quality attributes. Described here are the calculation methods, a discussion of the measure characteristics, and a brief example of using RobX to find a global optimal formulation across multiple quality characteristics within the framework of a mixture experimental design. The data presented are actual stability data generated during a formulation experimental design for a monoclonal antibody. We anticipate this approach will provide a good general measure of drug stability and will help to accelerate the drug development process. We believe it will improve formulation robustness, long-term stability, and the delivery of quality drug products to the patient. © 2011 Wiley Periodicals, Inc. and the American Pharmacists Association.

Diamond is used as detector material in high energy physics experiments due to its inherent radiation tolerance. The RD42 collaboration has measured the radiation tolerance of chemical vapour deposition (CVD) diamond against proton, pion, and neutron irradiation. Results of this study are summarized in this article. The radiation tolerance of diamond detectors can be further enhanced by using a 3D electrode geometry. We present preliminary results of a poly-crystalline CVD (pCVD) diamond detector with a 3D electrode geometry after irradiation and compare to planar devices of roughly the same thickness.

We describe the design of a ground based telescope which would be sensitive to extraterrestrial gamma rays with energies from 10 GeV to over 1000 GeV. Among other things, this telescope would enable the study of the emission mechanism of gamma rays from active galactic nuclei (AGNs), and of the interactions of gamma rays from AGNs with the intergalactic infrared radiation field (IIRF), possibly permitting an improved measurement of the Hubble constant. Calculations are presented which show that with the energy sensitivity and aperture of the telescope, as many as a thousand new sources of high energy gamma rays from outside of our Galaxy could be identified, about 10 of which would each be capable of measuring the Hubble constant to within 10% (assuming the parameters of the IIRF to be known).

The In Situ Storage Image Sensor (ISIS) is a monolithic active pixel sensor with memory cells in each pixel. The memory cells are implemented as a CCD register. A test device with parameters suitable for tracking charged particles has been constructed and characterized. The characteristics of a proof-of-principle ISIS device are described and results from using the sensor as a tracking device for high energy electrons are presented.

The TeraPixel Active Calorimeter (TPAC) sensor is a novel Monolithic Active Pixel Sensors (MAPS) device developed for use as the active layers of a large area, digital electromagnetic calorimeter (DECAL) at a future e+e− collider. Further applications, which include the tracking and vertex systems for future lepton colliders and LHC upgrades have been proposed and it is therefore essential to characterise the behaviour of the sensor for these applications. We present the first studies of radiation hardness testing of the TPAC sensor. The performance of the sensor has been evaluated after exposures up to 5 Mrad of 50 keV x-rays. Under realistic ILC operating conditions a maximum decrease in the signal to noise ratio of 8% (15%) was observed after 200 krad (5 Mrad) which is already sufficient for proposed applications in future e+e− colliders.

English editions of Euclid's Elements clashed over the arithmetization of mathematics. The editions of Henry Billingsley, Claude Dechales, and Isaac Barrow from the 16th and 17th centuries paid relatively little attention to mathematical primacy. In the 18th century, however, William Whiston asserted that algebraic representations, demonstrations, and proportions were more convenient than, yet as rigorous as, geometry. John Playfair's algebra adapted the Elements to modern audiences. Edmund Scarburgh, John Keill, Edmund Stone, and Robert Simson, however, attacked editions for alterations. Some editors tried to recover Euclid's original text. Most passages on this issue appeared in the editors' prefaces, Book II, Book V, or their notes. Copyright 2000 Academic Press. Des éditions anglaises des Éléments d'Euclid ne s'accordaient pas sur l'arithmétization de mathématiques. Les éditions d'Henry Billingsley, Claude Dechales, et Isaac Barrow des seizième et dix-septième siècles n'ont guère fait attention à la primatie mathématique. Cependant, au dix-huitième siècle, William Whiston a affirmé que les représentations, démonstrations, et proportions algébriques étaient plus commodes que, pourtant aussi rigoureuses que, la géométrie. L'algèbre de John Playfair a ajusté les Éléments aux lecteurs modernes. Edmund Scarburgh, John Keill, Edmund Stone, et Robert Simson cependant, ont attaqué les éditions pour les modifications. Quelques éditeurs ont essayé de retrouver le texte originel d'Euclid. La plupart des passages sur cette question se sont présentées dans les préfaces des éditeurs, Livre II, Livre V, ou leurs notes. Copyright 2000 Academic Press. MSC 1991 subject classifications: 01A20, 01A40, 01A45, 01A50

Abstract The Intermediate Silicon Layers (ISL) detector is part of the CDF upgrade for Run II. The ISL is a large radius (20–28 cm) double-side silicon tracker with a total active area of ≃3.5 m 2 . The full procedure for module production and electrical tests is described.

Revaccination with standard calf lymph vaccine was performed on 26 children who had received a primary vaccination with an attenuated smallpox vaccine, CVI-78, and 22 children who had received primary vaccination with standard calf lymph. Revaccination resulted in a vesicular reaction in 96 percent of those who had been vaccinated previously with CVI-78 and 73 percent of those vaccinated previously with standard calf lymph. All children had a positive hemagglutination-inhibition (HI) antibody titer either after primary vaccination or revaccination. Only 65 percent of those initially vaccinated with CVI-78 vaccine had positive neutralizing antibodies after revaccination. All children who received primary vaccination with standard calf lymph had postrevaccination neutralizing antibodies. The children who had neither a dermal nor a serologic response after primary vaccination responded as primary vaccinees on challenge with standard calf lymph.

The Linear Collider Flavour Identification (LCFI) Collaboration is developing the sensors, readout electronics and mechanical support structures for the vertex detector of the International Linear Collider (ILC). High speed readout is needed to ensure that the occupancy due to the pair production background at the ILC is kept below the 1% level. In order to satisfy this requirement, Column Parallel CCDs (CPCCDs), Column Parallel Readout chips (CPRs) and Column Parallel Driver chips (CPDs) have been developed. The CPCCD has to operate at a clock frequency of 50 MHz, which represents a difficult technical challenge due to the large sensor capacitance. The design and performance of the second generation CPCCD sensors, CPC2, and the new driver chip, CPD1, which meet these challenging requirements, are described.

The LCFI Collaboration is developing the sensors, readout electronics and mechanical support structures for the vertex detector (VXD) of the International Linear Collider (ILC), as well as studying the physics performance that the VXD will achieve. Since the VXD must provide excellent spatial resolution, the sensors must have very low mass and their power consumption must be small to ensure that the sensors themselves cause as little multiple scattering as possible and that gas cooling can be used to operate them at temperatures of about −40 C. High-speed readout is also needed to ensure that the occupancy due to the pair production background at the ILC is kept below the 1% level. In order to satisfy these strict requirements, Column Parallel CCDs (CPCCDs) and Column Parallel Readout chips (CPRs) have been developed. The readout chips must be able to operate synchronously with the CPCCDs and ensure fast signal processing with low noise and data compression. The design and performance of the latest version of the readout chip, the CPR2A, are described here.

An isoelectric focusing method (IEF) has been used to assess the charge heterogeneity profile of a monoclonal antibody during the early stages of product development. A more precise and sensitive ion exchange chromatography (IEC/CEX) method was developed and implemented as development progressed and was used concurrently with IEF for lot release and to monitor charge heterogeneity. Charge variants resolved by both methods (IEC and IEF) were purified and characterized. Tryptic peptide mapping and N- linked oligosaccharide profile analyses of the IEC and IEF fractions indicated a structural correlation between the charge variants separated by these two methods. The major sources of molecular heterogeneity were due to the variation in the sialyated carbohydrate structure and heavy chain C-terminal lysine truncation. By monitoring the rates of change in the charge heterogeneity profiles of the monoclonal antibody stored at elevated temperatures by the IEC and IEF methods, a positive correlation between the two methods was established. This approach enabled replacement of the IEF method with the more precise IEC method.

Abstract Both PD-1 and ILT4 (LILRB2, CD85d) have well established immunosuppressive effects that enable tumors to evade anti-tumor immunity. Tumor expression of PD-L1 and ILT4 has been associated with poor clinical outcomes in several cancer types. Recently, ILT4 activation has been postulated as a resistance mechanism for PD-1/PD-L1 blockade prompting the investigation of dual blockade of these pathways. Siu, L.L. et al. (2022 Clin Cancer Res 28: 57-70) reported encouraging results of the initial clinical study combining the ILT4 antagonist mAb MK-4830 with pembrolizumab. The combination was generally well tolerated and led to several durable responses including in PD-1/PD-L1 refractory patients. We have combined novel PD-1 (mAb E1A9) and ILT4 (mAb 7B1) antagonist antibodies into a tetravalent IgG-scFv format bispecific antibody (bsAb, CDX-585). CDX-585 was engineered to avoid effector functionality by eliminating interaction with Fcγ receptors, and to increase its half-life by enhancing binding to FcRn. As expected, CDX-585 inhibits interaction of PD-1 and ILT4 with their ligands, results in potent inhibition of PD-1 signaling and enhances myeloid cell inflammatory responses to stimulation through toll like receptors (LPS) or CD40 (CD40 agonist mAb). In mixed lymphocyte reactions CDX-585 promoted greater T cell activation than the combination of the parental mAbs. Similarly, CDX-585 demonstrated superior anti-tumor activity over the combination of the parental mAbs in a humanized mouse model of melanoma. CDX-585 has completed GMP manufacturing and IND-enabling activities. Doses of CDX-585 up to 60 mg/kg were well tolerated in cynomolgus macaques and displayed a favorable pharmacokinetic profile. Together these data demonstrate that CDX-585 effectively combines PD-1 and ILT4 blockade into one molecule with favorable biophysical and functional characteristics supporting the initiation of a dose-escalation clinical trial in patients with advanced solid tumors. Citation Format: Lawrence J. Thomas, Laura A. Vitale, Michael Murphy, Collin Xia, Zeyu Peng, Asma Ejas, Montu Patel, James Boyer, April R. Baronas, Thomas O'Neill, Jenifer Widger, Laura Mills-Chen, Andrea Crocker, Mark Ma, Mingjiu Chen, Hugh M. Davis, Russ A. Hammond, Cherie Taglienti, Michael Yellin, Joel Goldstein, Diego Alvarado, Henry C. Marsh, Tibor Keler. CDX-585, a novel bispecific antibody targeting PD-1 and ILT4 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2963.

Abstract Rationale: Reduction of mast cells (MCs) through stem cell factor (SCF) neutralization combined with inhibition of the alarmin TSLP in a single molecule may result in broader efficacy in inflammatory disorders. Methods: Monoclonal antibodies (mAb) separately targeting SCF and TSLP were generated from immunized mice. A potent neutralizing antibody against each target was selected and humanized. Bispecific tetravalent antibodies (bsAbs) with specificity towards SCF and TSLP were generated. Candidate bsAbs were tested in vitro for neutralizing activity and evaluated in pilot non-human primates (NHP) studies. Results: We discovered a novel anti-SCF neutralizing mAb (SCF-12) that potently inhibits SCF-dependent KIT phosphorylation and SCF-enhanced degranulation of primary human MCs. Separately, a novel anti-TSLP mAb (1D10) inhibits TSLP receptor binding and TSLP-dependent TARC release from human monocyte-derived DCs. bsAbs combining humanized SCF-12 and 1D10 with good expression, biophysical profile and robust neutralizing activity against each target were generated. Following demonstration of MC depletion in NHPs with SCF-12, we conducted a pilot NHP study with two lead candidate bsAbs. Pharmacokinetic, pharmacodynamic and tolerability data from this study will be presented. Conclusions: SCF x TSLP bsAbs were successfully generated from antibodies that neutralize soluble SCF, leading to MC reduction in NHP, and TSLP, a key alarmin implicated in inflammatory and autoimmune disorders. Combined inhibition of these two pathways using a single molecule may be of broad utility in multiple inflammatory disorders. Celldex Therapeutics

A size exclusion HPLC method has been developed to determine the protein concentration of pharmaceutical formulations of recombinant acidic fibroblast growth factor (aFGF). These topical aFGF formulations not only contain low levels of protein mass (50 μg ml−1), but also include buffer ions, polysaccharide polyanions to conformationally stabilize aFGF and 1% hydroxyethylcellulose to increase the solution's viscosity. A cesium chloride mobile phase is utilized during SEC-HPLC to dissociate aFGF from the pharmaceutical excipients and to minimize nonspecific interaction of the protein with the column matrix. The protein content of a viscous aFGF formulation is determined by comparison of aFGF peak areas to standards of known concentration. Fluorescence spectroscopy was utilized to directly demonstrate that the protein remains in its native conformation during sample preparation and analysis.

BACKGROUND: Only one surface cysteine residue (285) has been thought to be involved in D antigenicity, according to studies using lyophilized or nonlyophilized red cell membranes. However, it has been reported that a 17‐kDa chymotryptic fragment containing the N‐terminus but not this cysteine residue is associated with D antigenicity. STUDY DESIGN AND METHODS : The role of the sulfhydryl (SH) group in D, c, and E antigenicity is assessed by using intact red cells treated with the reagents N‐ethylmaleimide, 5,5′‐dithiobis(2‐nitrobenzoic acid), and 2‐ (4′‐maleimidylanilino)‐ naphthalene‐6‐sulfonic acid. Antigenicity was appraised by hemagglutination titers and immunoprecipitation using human anti‐D, ‐c, and ‐E. RESULTS : Treatment with N‐ethylmaleimide or 5,5′‐dithiobis(2‐nitrobenzoic acid) at various concentrations (&lt; or = 5 mM) or for various times (&lt; or = 120 min) did not cause significant decrease in hemagglutination titers as compared to untreated intact red cells. Moreover, immunoprecipitation of Rh antigen‐carrying peptides by human anti‐D was not affected by prior treatment with N‐ethylmaleimide or 2‐(4′maleimidylanilino)‐naphthalene‐6‐sulfonic acid. Efficacy of blockage of SH groups was demonstrated by inhibition of palmitic acid uptake by the Rh polypeptides for prior treatment with N‐ethylmaleimide and by the presence of fluorescent Rh polypeptides for prior treatment with 2‐(4′maleimidylanilino)‐naphthalene‐6‐sulfonic acid. CONCLUSION : SH group involvement is not essential for D, c, or E antigenic expression in intact red cells.

The Intermediate Silicon Layers detector (ISL) is a large radius silicon tracker, installed in the CDF detector for the RUN II of the Tevatron Collider. With almost 4 m2 of double-sided silicon sensors and 300,000 electronic channels it represents the biggest system of this kind ever built. The construction and installation phases, the performed quality assurance tests as well as the problems encountered are reviewed. RUN II of the Tevatron officially started on March 1st, 2001. Although the CDF silicon system is still being commissioned, results on the performance of the ISL detector obtained using the first data are presented.

The innermost layer (L00) of the Run IIa silicon detector of CDF was planned to be replaced for the high luminosity Tevatron upgrade of Run IIb. This new silicon layer (L0) is designed to be a radiation tolerant replacement for the otherwise very similar L00 from Run IIa. The data are read out via long, fine-pitch, low-mass cables allowing the hybrids with the chips to sit at higher z(/spl sim/70 cm), outside of the tracking volume. The design and first results from the prototyping phase are presented. Special focus is placed on the amount and the structure of induced noise as well as signal-to-noise values.

Abstract The LCFI Collaboration is developing the sensors, readout electronics and mechanical support structures for the Vertex Detector (VXD) of the International Linear Collider (ILC). High-speed readout is needed to ensure that the occupancy due to the pair production background at the ILC is kept below 1% level. In order to satisfy this requirement, Column Parallel CCDs (CPCCDs) and Column Parallel Readout chips (CPRs) have been developed. The CPCCD has to operate at a clock frequency of 50 MHz, which represents a difficult technical challenge due to the relatively large sensor capacitance. The design and performance of planar transformers, which can be used to provide the required 20 A clock current, are described.

The Linear Collider Flavour Identification (LCFI) collaboration has successfully developed the first prototype of a novel particle detector, the In-situ Storage Image Sensor (ISIS). This device ideally suits the challenging requirements for the vertex detector at the future International Linear Collider (ILC), combining the charge storing capabilities of the Charge-Coupled Devices (CCD) with readout commonly used in CMOS imagers. The ISIS avoids the need for high-speed readout and offers low power operation combined with low noise, high immunity to electromagnetic interference and increased radiation hardness compared to typical CCDs. The ISIS is one of the most promising detector technologies for vertexing at the ILC. In this paper we describe the measurements on the charge-shielding properties of the p-well, which is used to protect the storage register from parasitic charge collection and is at the core of device's operation. We show that the p-well can suppress the parasitic charge collection by almost two orders of magnitude, satisfying the requirements for the application.

Abstract The TAM receptors (Tyro3/Axl/MerTK) family of receptor tyrosine kinases (RTKs) are important negative regulators of innate immunity. TAM receptor activation in myeloid cells by its ligands Gas6 or Protein S (PROS) promotes phosphatidylserine-dependent efferocytosis of apoptotic cells, inducing a tolerogenic state and mediating resolution of inflammation. TAM-deficient mice exhibit phenotypes consistent with systemic inflammation and autoimmunity. Importantly, individual ablation of TAM receptors can confer tumor immunity, increased pro-inflammatory cytokines and tumor lymphocyte infiltration, leading to the proposal that TAM receptors act as checkpoints of innate immunity. We hypothesize that pharmacological targeting of this family of receptors with monoclonal antibodies (mAbs) may lead to a similar pro-inflammatory response and recapitulate the antitumor effects observed in TAM-deficient mice. From a panel of human anti-MerTK, Axl or Tyro3 mAbs derived from phage-display libraries or human IgG expressing mice we identified unique mAbs that markedly enhanced cytokine and chemokine release from primary human immune cells, alone or in the presence of inflammatory stimuli. Interestingly, the qualitative and quantitative pattern of cytokine response from dendritic cells was very similar using antibodies targeting the individual TAM receptors, and also similar to activation of dendritic cells using a CD40 agonist mAb, suggesting that TAM-targeting mAbs can promote immune activation. We identified surrogate mAbs targeting mouse TAM receptors that elicited similar responses in vivo, and demonstrated antitumor activity when dosed alone, or in combination with PD-1/L1 blockade in syngeneic tumor models. In addition, human MerTK transgenic, and TAM knockout mice have been generated and characterized in order to establish in vivo proof-of-concept with human TAM mAbs. Overall, pharmacological modulation of TAM receptors with mAbs enhances cytokine production in human and murine model systems consistent with the published role of TAMs as negative regulators of innate immunity. Mechanistic and proof-of-concept studies support further development of these mAbs as novel approaches to overcome these checkpoints of the innate immune response. Citation Format: Diego Alvarado, Laura Vitale, Mike Murphy, Thomas O'Neill, Andrew Proffitt, Jay Lillquist, Gwenda Ligon, Komal Patel, Anna Wasiuk, Jeff Weidlick, Jenifer Widger, Laura Mills-Chen, Andrea Crocker, Colleen Patterson, Russell A. Hammond, Li-Zhen He, Joel Goldstein, Lawrence J. Thomas, Henry C. Marsh, Tibor Keler, Richard Gedrich. Monoclonal antibodies targeting the TAM family of receptor tyrosine kinases [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1555.

Abstract CD40 is a key molecule in the regulation of immune responses and its activity can be modulated using antibodies. In particular, agonist CD40 antibodies are highly effective in preclinical tumor models either through direct interaction with CD40-expressing lymphomas, or indirectly through the activation of an adaptive anti-tumor immune response. To date, limited clinical data have been reported with strong CD40 agonist antibodies; nonetheless it seems likely that targeting this pathway will require a balance between the benefits of immune stimulation to drive anti-tumor responses, and the damage that can result from non-specific immune cell activation. We set out to develop novel human anti-CD40 antibodies with different levels of agonist activity to identify a lead candidate for systemic application. Anti-CD40 monoclonal antibodies (mAbs) were generated by immunization of human Ig transgenic mice with recombinant and cell surface expressed human CD40. Hybridomas developed from these mice were screened using CD40 binding assays and activity on a reporter cell line engineered to express CD40 and NFêB-responsive luciferase. The variable regions of lead antibodies that displayed differential activity were cloned into vectors containing human IgG1 or IgG2 constant domains and expressed in CHO cells. These human CD40 mAbs were further characterized by analysis of binding affinity, CD40L blocking activity, B cell and dendritic cell activation, and anti-tumor activity in xenograft tumor models. We found a wide range of activities among the CD40 mAbs that is linked to epitope specificity as well as the isotype. In general, the IgG2 isotype mAbs had greater signaling activity than their IgG1 counterparts. The lead candidate mAbs are undergoing additional testing related to functional and toxicity parameters before a final candidate is nominated for clinical development. Citation Format: Laura A. Vitale, Thomas O’Neill, Jenifer Widger, Andrea Crocker, Li-Zhen He, Jeffrey Weidlick, Karuna Sundarapandiyan, James Storey, Lawrence Thomas, Joel Goldstein, Henry C. Marsh, Tibor Keler. Development and characterization of novel CD40 antibody agonists for cancer immunotherapy. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4866.

We have tested the first 4T Monolithic Active Pixel Sensor (MAPS) for particle physics, FORTIS in a beam test. We have measured a signal-to-noise ratio of more than 100 for MIPs due to the excellent noise performance of the 4T architecture. Two versions of the sensor were tested; with and without deep P-well areas in-pixel. The deep P-well areas allow the incorporation of PMOS transistors inside the pixels without signal charge loss. The measured position resolutions were around 2 μm.

The production of a Standard Model Higgs boson in association with a top quark pair at the upcoming high luminosity run (15 fb -1 integreted luminosity) of the Fermilab Tevatron [Formula: see text] is revisited. For Higgs masses below 140 GeV we demonstrate that the production cross section times branching ratio for [Formula: see text] decays yields a significant number of events and that this mode is competitive with and complementary to the searches using [Formula: see text], ZH associated production. For higher mass Higgs bosons the H → W + W - decays are more difficult but have the potential to provide a few spectacular events.