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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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Enabling Epigenetics Studies from HTS to SAR : A Novel HTRF® Platform to Identify and Characterize Reader Domain Inhibitors
T. Roux1, M. Badol1, N. Douayry1, L. Sergeant1, E.Trinquet1, F. Degorce1, S. Milhas2, S. Betzi2, C. Derviaux2, C. Eydoux3, J. Letienne2, A. Lugari2, Y. Collette2, J-C. Guillemot3 et X. Morelli2

Discover a novel HTRF platform to identify and characterize the vast variety of epigenetic binding domain.

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Scientific News
Natural Protein Points to New Inflammation Treatment
Findings may offer insight to effective treatments for inflammatory diseases, such as rheumatoid arthritis, psoriasis, and multiple sclerosis.
Tricked-Out Immune Cells Could Attack Cancer
New cell-engineering technique may lead to precision immunotherapies.
Therapy Halts Progression of Lou Gehrig’s Disease
Researchers at Oregon State University announced today that they have essentially stopped the progression of amyotrophic lateral sclerosis (ALS), or Lou Gehrig’s disease, for nearly two years in one type of mouse model used to study the disease – allowing the mice to approach their normal lifespan.
Crouching Protein, Hidden Enzyme
A new study led by scientists at The Scripps Research Institute (TSRI) and the University of California (UC), Berkeley shows how a crucial molecular enzyme starts in a tucked-in somersault position and flips out when it encounters the right target.
Utilizing Antibodies from Ebola Survivors
A collaborative team from The University of Texas Medical Branch at Galveston, Vanderbilt University, The Scripps Research Institute and Integral Molecular Inc. have learned that antibodies in the blood of people who have survived a strain of the Ebola virus can kill various types of Ebola.
Engineering Foe into Friend
Bose Grant awardee Jacquin Niles aims to repurpose the malaria parasite for drug delivery.
Important Regulator of Immune System Decoded
Plasma cells play a key role in our immune system. Now scientists at the Research Institute of Molecular Pathology (IMP) in Vienna, Austria, and at the Walter and Eliza Hall Institute (WEHI) in Melbourne, Australia, succeeded in characterizing a central regulator of plasma cell function.
Bacterial Superglue for Faster Vaccine Development
An interdisciplinary team of Oxford University researchers has devised a new technique to speed up the development of novel vaccines.
Enumeral, MD Anderson Enter Into Collaboration
Strategic collaboration aims to discover and develop potentially novel antibodies against specified targets for immunotherapy.
Joint venture for GlycoMar Limited and MicroA AS
Scottish biotechnology company GlycoMar and Norwegian technology company MicroA are pleased to announce their joint venture business Prasinotech Ltd. The new company is registered in Scotland.
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