|A Simple Plate Based Assay Using pH Sensor Dye to Screen for Internalizing Antibody|
Nidhi Nath, Becky Godat, Cesear Corona, Chad Zimprich, Mark McDougall, Poncho Meisenheimer, Marjeta Urh
Receptor mediated internalization is a key mechanism of action (MOA) for antibody drug conjugates (ADCs). However, current methods of studying antibody internalization have several limitations including: 1) A multistep process not suitable for screening; 2) Low signal-to-background ratios; 3) Not suitable for kinetic measurements. We have developed a method that mitigates problems associated with traditional internalization assays.
|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.
|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.
|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.
|Modelling CLL cell and T-cell Migration in a Dynamic Circulating Model of CLL|
Elisabeth Walsbyl, Paul Brennan', Guy Pratte, Andrea Buggins3, Tanja N Hartmann'', Chris Fegan1 and Chris Pepper'
We have recently developed a novel circulating model of chronic lymphocytic leukaemia (CLL) that mimics the transient interactions that take place between circulating lymphocytes and vascular endothelium. Here we show that both normal and malignant lymphocytes actively underwent transendothelial migration.
|Quantification of Natural Killer Cell-Mediated Cytotixicity using Celigo Imaging Cytometry|
Leo L. Chan, Srinivas S. Somanchi, Kelsey Rosbach, Dean A. Lee
Time-course tracking of % lysis eliminates multiple controls & the effect of non-uniform cell seeding in the final cytotoxicity calculation. The # of cells used is less than the cells needed for Release assays & Flow Cytometry. Flow cytometry & Release assays require a seeding density of 100K target cells increasing the number of effector cells to the millions. The visual observation of ADCC or CDC on the images can be convincing to conclude the functionality of antibodies or complements.
|Non-invasive Liquid Biopsy|
Frauke Henjes1, Lise Aagaard1, Darren Ellis1, Hannah Lindstroem1, Morten Oksvold2,3, Axl Neurauter1, Ketil W. Pedersen1.
Paramagnetic Dynabeads® magnetic beads provide a versatile, automation friendly tool for fast sensitive and high throughput isolation that can be tailored to isolate specific circulating biomarkers such as cells, exosomes and nucleic acids.
|Novel culture medium using a small-molecule agonist of thrombopoetin receptor|
Hondo M1; Nishino T2; Inamura M1
ReproHSC medium for the culture of hematopoietic stem cells. facilitates the culture and expansion of CD34+/CD38- cells that retain their HSC properties
|Flexible automated platform for blood group genotyping on DNA microarrays|
S. Paris1, D. Rigal1, V. Barlet1, M. Verdier1, N. Coudurier2, P. Bailly3, J.-C. Brès1,4
The purpose of this project was to set up and validate a flexible robotic platform using 96-well DNA microarray for multiplex blood group genotyping.
|THE MASTOCYTOSIS SOCIETY SURVEY ON MAST CELL DISORDERS: Part 2-Clinical Experiences, Co-Morbidities, Diet, Families and Opinions|
aNancyRussell, DrPH, aSusanJennings, PhD, aBlairJennings, BS, aValerie Slee , RN, BSN, aLisa Sterling, BS, bMariana Castells, MD, PhD, FAAAAI,cPeter Valent, MD, bCem Akin, MD, PhD, FAAAAI
Mast cell diseases such as mastocytosis and mast cell activation syndromes involve abnormal proliferation or activation of these cells leading to many potentially debilitating symptoms. In order to determine the characteristics and experiences of people known or suspected to have a mast cell disorder, The Mastocytosis Society (TMS), a U.S. based patient advocacy, research and education organization, conducted a survey of patients.
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