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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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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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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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Automated in-gel digestion on a commercial autosampler directly coupled to nanoLC-MS/MS
Achermann François, Bolliger Reto, Buchs Natasha, Doiron Nicholas, Lagache Braga Sophie, Heller Manfred, Boehm Guenter

SDS-PAGE separates protein samples from LC-MS incompatible contaminations, and is frequently used to fractionate proteins of entire proteomes. One disadvantage is that gel lanes have to be cut into many slices, followed by in-gel digestion of proteins and extraction of peptides. The number of these gel slices goes into the hundreds, rendering this process very repetitive and prone to mistakes and errors during sample handling. Automation reduces such risks and improves reproducibility.

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Automated sample preparation workflows for quantitative proteomics applications
Oliver Popp1, Lucas Luethy2, Tamara Kanashova1, HaAn Nguyen1, Julia Kikuchi1, Guenter Boehm2, Thomas Blenkers3, Andreas Bruchmann3, Gunnar Dittmar1

Mass spectrometry based proteomics requires large scale identification of peptides, and depends upon efficient sample preparation. Recently, we presented two automated protein-digestion setups, in-solution and in-gel digestion. We extended these techniques by implementing dimethyl labelling (DML). Furthermore, we established an automated phospho-peptide (PP) enrichment procedure in a 96-well formate, generating phospho-proteomic data in very short time.

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Characterization of Protein and Protein Aggregates using Temperature-controlled Hollow-Fiber Flow-FFF
Trevor Havard, Florian Meier, Evelin Moldenhauer, Soheyl Tadjiki, Thorsten Klein

Reproducibilty Improvements in Field Flow Fractionation can be achieved on two fronts. Instrument design and control, the system used in for this poster is does not require flow controllers or switching valves and there for produces the same conditions in every case. Fractionater design, the design of the cartridges used in this poster maintain excellent conditions to maintain constant pressure at the membrane removing unwanted effects of sale relaxation above the membrane s

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Specificity of highly potent miRNA inhibitors
Barbara Roberston, Andrew Dalby, Yuriy Fedorov, Jon Karpilow, Anastasia Khvorova, Devin Leake, Annaleen Vermeulen

Specificity of highly potent miRNA inhibitors

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HOMOGENEOUS ADCs BEARING TWO DIFFERENT PAYLOADS SHOW STRONG SYNERGY IN TUMOR KILLING
Lisha Allen, Yi Mi, Muhammad Abbas, Wolfgang Richter, and Sean Hu

We presented a one-pot reaction to prepare site-specific ADCs using available mABs on hand without re-engineering. We have identified a novel non-cleavable linker, which increases not only ADC stability, but also potency. We also demonstrate synergy in cancer cell killing of hybrid ADC loaded with two different toxins

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Toward a Comprehensive Bottom-up and Top-down Analysis of the NIST Reference Monoclonal Antibody
Chris Becker1, Eric Carlson1, Wilfred Tang1, Yong Kil1, Marshall Bern1, John Schiel2, Lisa Kilpatrick2, Trina Formolo2

Development of ADC and other therapeutic proteins require careful characterization of sequence, post-translational modifications, sequence variants, and degradants. In this study, a NIST mAb interim reference material was analyzed by new bioinformatics tools (Byonic™, Byologic®, and Byomap™) allowing one to efficiently identify and quantify modifications down to trace levels.

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Scientific News
ASMS 2016: Targeting Mass Spectrometry Tools for the Masses
The expanding application range of MS in life sciences, food, energy, and health sciences research was highlighted at this year's ASMS meeting in San Antonio, Texas.
Long-Term Culturing of Adult Stem Cells
A new procedure developed by Harvard Stem Cell Institute researchers (HSCI) at Massachusetts General Hospital (MGH) may revolutionize the culturing of adult stem cells.
Solutions for Biotherapeutic Characterization
Innovation to speed the routine.
Plant-Based Vaccine Among Front Runners In Search For New Polio Jab
A researcher from Norwich is part of a consortium that has been awarded $1.5 million to develop safer polio vaccines, using a new technique developed at the John Innes Centre.
Harnessing Helpful Microbes
Seeking to further harness microbes’ many uses, the federal government has launched the National Microbiome Initiative (NMI) to “foster the integrated study of microbiomes across different ecosystems.”
What Makes a Good Scientist?
It’s the journey, not just the destination that counts as a scientist when conducting research.
A New Approach to Chemical Synthesis
Communesins, originally found in fungus, could hold potential as cancer drugs.
Better Animal Model to Improve HIV Vaccine Development
Penn study identifies a new tool to produce better HIV vaccine designs.
First Large-Scale Proteogenomic Study of Breast Cancer
The study offers understanding of potential therapeutic targets.
Developing a More Precise Seasonal Flu Vaccine
During the 2014-15 flu season, the poor match between the virus used to make the world’s vaccine stocks and the circulating seasonal virus yielded a vaccine that was less than 20 percent effective.
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