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An IgG Cleaving Protease from S equi with Improved Activity Against Mouse IgGs
Chris Hosfield, Philip Compton, Luca Fornelli, Paul Thomas, Neil L. Kelleher, Michael Rosenblatt & Marjeta Urh

Here we have expressed and purified a modified recombinant IdeZ, and show that it has significantly improved activity against mouse IgG2a and IgG3 subclasses when compared to IdeS. We also demonstrate the use of IdeZ in LC-MS workflows for human and mouse IgG characterization.

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Better Cell-Based Assays for Anti-CTLA-4, Anti-PD-1/PD-L1, and Bispecific Immunotherapy Drug Studies
Richard Somberg, Mei Cong, Pete Stecha, Natasha Karassina, Jim Hartnett, Zhi-Jie Jey Cheng, and Frank Fan

Here we report the development of a panel of robust reporter assays to measure the potencies for biologics in immunotherapy. These assays reflect mode of action and can serve as valuable tools in immunotherapy drug development and discovery.

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CellTiter-Glo® 2.0: A Novel Luminescent Cell Viability Assay with Greatly Enhanced Storage Stability
Michael P. Valley, James Unch, Poncho L. Meisenheimer, James J. Cali, and Dan F. Lazar

Here we report on the attributes of a novel ATP detection reagent for cell viability with all of the assay performance of the previous CellTiter-Glo® Reagent, but now with markedly enhanced stability as a single component in a liquid format. These new features provide for much greater ease-of-use in that storage of the reagent at 4°C eliminates the requirement for reagent thawing and minimizes temperature equilibration time.

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Design and Validation of Bioluminescent Assays for 3D Cell Culture Models Poster
Terry L. Riss, Michael P. Valley, Chad A. Zimprich, Andrew L. Niles, Kevin R. Kupcho and Dan F. Lazar

Cells cultured in 3D model systems often acquire relatively large in vivo-like structures compared to the thickness of a 2D monolayer of cells grown on standard plastic plates.

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Development of a Robust Reporter-based T cell Activation Assay for Therapeutic Biologics in Immunotherapy
Zhi-jie Jey Cheng, Pete Stecha, Jim Hartnett, Frank Fan, and Mei Cong

Jurkat T-cells stably expressing luciferase reporter driven by IL2 promoter or NFAT-RE, are used as effector cells. Tumor cell lines endogenously expressing cancer antigen are used as antigen presenting cells (APC). By co-cultivating the two cell lines in the presence of CD3 bispecific antibody, TCR/CD3 is activated in Jurkat effector cells. Luciferase activity is up regulated through IL-2 promoter or NFAT-RE activation.

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iPSC-Derived Cardiomyocytes and Luciferase Reporters: A Robust Reporting Platform for Monitoring Cardioprotection and Pathway Biology in Endogenous Human Tissue Cells
Fiene, S., Thompson, A., Niles, A., Robers, M., Anson, B.

Pathophysiological conditions, medical interventions, and off-target toxicities can all result in cellular oxidative stress. In cardiac myocytes, prolonged and/or excessive oxidative stress can lead to cardiotoxicity: a primary cause of developmental delays, black-box warnings, and post-launch withdrawal of pharmaceuticals.

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Reporter Bioassays to Assess Therapeutic Antibodies for Immunotherapy Programs
Mei Cong, Zhi-Jie Jey Cheng, Pete Stecha, Jun Wang, Jamison Grailer, Natasha Karassina, Jim Hartnett, and Frank Fan

Immunotherapy, also called biologic therapy or biotherapy, stimulates certain parts of the immune system to fight diseases such as cancer. Important drug targets in immunotherapy include: Co-inhibitory receptors, such as PD-1/PD-L1, CTLA-4, LAG3, Tim3; and co-stimulatory receptors, such as GITR, CD40, OX40, 4-1BB.
Current approaches to assaying these targets are cumbersome and variable. Here we offer an improved in vitro bioassay approach.

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ROS-Glo™ H2O2 Assay: A Luminescent Assay for Detection of Reactive Oxygen Species Poster
Gediminas Vidugiris, Sarah Duellman, John Shultz, Jolanta Vidugiriene, Hui Wang, Jean Osterman, Wenhui Zhou, Poncho Meisenheimer and James J. Cali

H2O2 is a reactive oxygen species (ROS) that is measured in cells as a marker of oxidative stress. It is also measured as a marker of enzyme activities that either consume or produce H2O2. It is desirable to screen chemical compounds for their capacity to alter H2O2 levels in cultured cells or for their effects on H2O2 levels in enzyme reactions.

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Testing a Novel Real Time Cell Viability Assay
Amy Landreman, Sarah Duellman, Wenhui Zhou, Jolanta Vidugiriene, Brad Hook

Recently developed assay technologies make it possible to use multi-well plate readers to measure the number of live or dead cells in culture in real time over a period of days. Live cells are measured in real time by adding a reagent containing a shrimp-derived luciferase and a pro-substrate directly to the culture medium. Only viable cells can convert the pro-substrate into a luciferase substrate and generate light.

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Scientific News
Flowering Regulation Mechanism Discovered
Monash researchers have discovered a new mechanism that enables plants to regulate their flowering in response to raised temperatures.
Turning Skin Cells into Heart, Brain Cells
In a major breakthrough, scientists at the Gladstone Institutes transformed skin cells into heart cells and brain cells using a combination of chemicals.
Nanoparticles Present Sustainable Way to Grow Food Crops
Nanoparticle technology can help reduce the need for fertilizer, creating a more sustainable way to grow crops such as mung beans.
How Scientists Use DNA to Track Disease Outbreaks
They’re the top questions on everyone’s mind when a new disease outbreak happens: where did the virus come from? When did this happen? How long has it been spreading in a particular country or group of people?
Genetic Risk Factors of Disparate Diseases Share Similar Biological Underpinnings
Penn Institute for Biomedical Informatics and colleagues identify "roadmap" of disease mechanisms to identify candidate drug targets.
Drugs that May Combat Deadly Antibiotic-Resistant Bacteria Uncovered
Study identifies 79 compounds that inhibit carbapenem-resistant Enterobacteriaceae (CRE).
Stem Cells Know How to Unwind
Research led by the Babraham Institute with collaborators in the UK, Canada and Japan has revealed a new understanding of how an open genome structure supports the long-term and unrestricted developmental potential in embryonic stem cells.
HIV Particles Used to Trap Intact Mammalian Protein Complexes
Belgian scientists from VIB and UGent developed Virotrap, a viral particle sorting approach for purifying protein complexes under native conditions.
Childhood Asthma Research Receives $2M
Research into the impact of a child’s upbringing and social and physical environments on the development of asthma will receive $2 million to tackle the condition that affects as many as one in three Canadians.
Growing Stem Cells More Safely
Nurturing stem cells atop a bed of mouse cells works well, but is a non-starter for transplants to patients – Brown University scientists are developing a synthetic bed instead.
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