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5th Annual CAR-TCR Summit 2019

Visit us at Booth #25 to learn about new tools for studying Target Cell Killing, Endogenous Biology, Intracellular Protein Interactions and more!

  •  Seaport Hotel and World Trade Center, Boston, MA.
  •  Tuesday, September 10, 2019 – Friday, September 13, 2019

Come to the Promega booth and learn about:

Target Cell Killing
Cell Health Assays
Bioassays
Custom CRISPR Capabilities

Can’t make it to Boston? See what Promega has to offer for your research needs.

Explore Biologics Drug Discovery

Be sure to attend our Tech Slam and Scientific Poster Presentation:

Brock Binkowski

A Novel, Bioluminescent Assay for the Selective Detection of Target Cell Killing in Mixed Cultures
Presented by Brock Binkowski, PhD, Sr. Research Scientist

Abstract:

Efforts to develop and commercialize cellular immunotherapies would benefit from assays that selectively monitor target cell death that are sensitive and easy-to-use. To address this, we have developed an approach to selectively quantify target cell death using a gain-of-signal assay format and bioluminescence read-out. The method relies on the release of a HiBiT-tagged protein from target cells following cell lysis. HiBiT, an 11 a.a. peptide tag, binds to cell-impermeable Large BiT (LgBiT), a 17.6 kDa protein, to reconstitute NanoBiT Luciferase. Target cells are engineered to express a HiBiT-tagged protein using either ectopic expression or CRISPR/Cas9 to tag endogenous lactate dehydrogenase (LDH), and cell lysis is quantified by adding a detection reagent containing LgBiT and furimazine substrate (no medium removal). The signal is proportional to the amount of target cell death, and measurements can be made using endpoint or kinetic formats. Cell lines have low rates of spontaneous release and fusion proteins that are stable in the extracellular medium, enabling assays up to 24 hours or more. The bright signal from NanoBiT Luciferase allows the use of low numbers of target cells per well (e.g. 2,500), and the assay can detect very low levels of target cell death (e.g. <5%). We demonstrate this approach using T cell redirection mediated by a bispecific antibody or chimeric antigen receptor (CAR), together with ADCC mediated by PBMCs. The assay provides a sensitive, simple read-out to support discovery research and/or QC lot release applications.

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