Characterization of bispecific T-cell Engager (BiTE) antibodies with a high-capacity T-cell dependent cellular cytotoxicity (TDCC) assay.
| Autor: | Nazarian AA; Therapeutic Discovery, Amgen Inc., Thousand Oaks, CA, USA aaronn@amgen.com., Archibeque IL; Oncology Research, Amgen Inc., Thousand Oaks, CA, USA., Nguyen YH; Therapeutic Discovery, Amgen Inc., Thousand Oaks, CA, USA., Wang P; Therapeutic Discovery, Amgen Inc., Thousand Oaks, CA, USA., Sinclair AM; Oncology Research, Amgen Inc., Thousand Oaks, CA, USA., Powers DA; Therapeutic Discovery, Amgen Inc., Thousand Oaks, CA, USA. |
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| Jazyk: | angličtina |
| Zdroj: | Journal of biomolecular screening [J Biomol Screen] 2015 Apr; Vol. 20 (4), pp. 519-27. Date of Electronic Publication: 2014 Dec 04. |
| DOI: | 10.1177/1087057114561405 |
| Abstrakt: | The Bispecific T-cell Engager (BiTE) antibody modality is a clinically validated immunotherapeutic approach for targeting tumors. Using T-cell dependent cellular cytotoxicity (TDCC) assays, we measure the percentage of specific cytotoxicity induced when a BiTE molecule engages T-cells, redirects T-cell mediated cytolysis, and ultimately kills target cells. We establish a novel luminescence-based TDCC assay quantified by measuring cell viability via constitutive expression of luciferase. The luciferase-based TDCC assay performance is valid and comparable to an adenosine triphosphate (ATP)-based detection method. We demonstrate that the luciferase-based TDCC assay is an efficient homogeneous assay format that is amenable to both suspension and adherent target cells. The luciferase-based TDCC assay eliminates the need for plate-washing protocols, allowing for higher-throughput screening of BiTE antibodies and better data quality. Assay capacity is also improved by performing serial dilutions of BiTE antibodies in 384-well format with an automated liquid handler. We describe here a robust, homogeneous TDCC assay platform with capacity for in vitro assessment of BiTE antibody potency and efficacy using multiple tumor cell lines and T-cell donors. (© 2014 Society for Laboratory Automation and Screening.) |
| Databáze: | MEDLINE |
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