|A Synthetic CRISPR-Cas9 System for Homology-directed Repair|
John A. Schiel, Maren M. Gross, Emily M. Anderson*, Eldon T. Chou, Anja van Brabant Smith Dharmacon, part of GE Healthcare, 2650 Crescent Drive, Lafayette, CO 80026, USA
Synthetic, dual-RNA-encoded Cas9 is used for precise homology-directed repair (HDR) gene engineering. Both short and long (GFP) inserts are covered.
|Mobility of Aeroallergens in Home: Effect of Location of Air Sampling and Implication for Evaluation of Patient Exposure|
Julian Gordon1, Paul Detjen, Andrea Wachter & Prasanthi Gandhi1
The Inspirotec sampler permitted the easy testing of multiple locations within a household. Air sampling simultaneously at 12 locations by other technologies would have been technically challenging. These were run by an untrained operator.
|Building a digital pathology ecosystem for education and research|
Yves Sucaet, Silke Smeets, Stijn Piessens, Sabrina D'Haese, Chris Groven, Wim Waelput, Peter In't Veld
We wanted to build a core digital pathology infrastructure to support different use cases. Various images platforms needed to be accessible through a single access point, and support different user profiles. We wanted a scalable solution that would allow interaction between equipment from different research groups.
We built a centralized infrastructure that integrates a variety of imaging platforms, and now have an interconnected network of heterogeneous and scalable information silos.
|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.