1. Microscale screening of antibody libraries as maytansinoid antibody-drug conjugates.
- Author
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Catcott KC, McShea MA, Bialucha CU, Miller KL, Hicks SW, Saxena P, Gesner TG, Woldegiorgis M, Lewis ME, Bai C, Fleming MS, Ettenberg SA, Erickson HK, and Yoder NC
- Subjects
- Antibodies, Monoclonal, Humanized immunology, Antibodies, Neoplasm immunology, Cell Line, Tumor, Drug Screening Assays, Antitumor methods, Humans, Immunoconjugates immunology, Neoplasms immunology, Antibodies, Monoclonal, Humanized pharmacology, Antibodies, Neoplasm pharmacology, Immunoconjugates pharmacology, Maytansine pharmacology, Neoplasms drug therapy
- Abstract
Antibody-drug conjugates (ADCs) are of great interest as targeted cancer therapeutics. Preparation of ADCs for early stage screening is constrained by purification and biochemical analysis techniques that necessitate burdensome quantities of antibody. Here we describe a method, developed for the maytansinoid class of ADCs, enabling parallel conjugation of antibodies in 96-well format. The method utilizes ∼ 100 µg of antibody per well and requires <5 µg of ADC for characterization. We demonstrate the capabilities of this system using model antibodies. We also provide multiple examples applying this method to early-stage screening of maytansinoid ADCs. The method can greatly increase the throughput with which candidate ADCs can be screened in cell-based assays, and may be more generally applicable to high-throughput preparation and screening of different types of protein conjugates.
- Published
- 2016
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