title: Microscale screening of antibody libraries as maytansinoid antibody-drug conjugates.
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.
authors.affiliation: a ImmunoGen, Inc. , Waltham , MA.
authors.forename: Kalli C
authors.initials: KC
authors.lastname: Catcott
authors.affiliation: a ImmunoGen, Inc. , Waltham , MA.
authors.forename: Molly A
authors.initials: MA
authors.lastname: McShea
authors.affiliation: b Novartis Institutes for Biomedical Research , Cambridge , MA.
authors.forename: Carl Uli
authors.initials: CU
authors.lastname: Bialucha
authors.affiliation: c Novartis Institutes for Biomedical Research , Emeryville , CA.
authors.forename: Kathy L
authors.initials: KL
authors.lastname: Miller
authors.affiliation: a ImmunoGen, Inc. , Waltham , MA.
authors.forename: Stuart W
authors.initials: SW
authors.lastname: Hicks
authors.affiliation: b Novartis Institutes for Biomedical Research , Cambridge , MA.
authors.forename: Parmita
authors.initials: P
authors.lastname: Saxena
authors.affiliation: b Novartis Institutes for Biomedical Research , Cambridge , MA.
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Gesner
authors.affiliation: c Novartis Institutes for Biomedical Research , Emeryville , CA.
authors.forename: Mikias
authors.initials: M
authors.lastname: Woldegiorgis
authors.affiliation: a ImmunoGen, Inc. , Waltham , MA.
authors.forename: Megan E
authors.initials: ME
authors.lastname: Lewis
authors.affiliation: a ImmunoGen, Inc. , Waltham , MA.
authors.forename: Chen
authors.initials: C
authors.lastname: Bai
authors.affiliation: a ImmunoGen, Inc. , Waltham , MA.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Fleming
authors.affiliation: b Novartis Institutes for Biomedical Research , Cambridge , MA.
authors.forename: Seth A
authors.initials: SA
authors.lastname: Ettenberg
authors.affiliation: a ImmunoGen, Inc. , Waltham , MA.
authors.forename: Hans K
authors.initials: HK
authors.lastname: Erickson
authors.affiliation: a ImmunoGen, Inc. , Waltham , MA.
authors.forename: Nicholas C
authors.initials: NC
authors.lastname: Yoder
mesh_terms: D061067:Antibodies, Monoclonal, Humanized / Q000276:immunology / Q000494:pharmacology*; D000912:Antibodies, Neoplasm / Q000276:immunology / Q000494:pharmacology*; D045744:Cell Line, Tumor; D004354:Drug Screening Assays, Antitumor / Q000379:methods; D006801:Humans; D018796:Immunoconjugates / Q000276:immunology / Q000494:pharmacology*; D008453:Maytansine / Q000494:pharmacology*; D009369:Neoplasms / Q000188:drug therapy* / Q000276:immunology
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D000912:Antibodies, Neoplasm; D018796:Immunoconjugates; D008453:Maytansine
keywords: Antibody; antibody-drug conjugate; conjugation; high-throughput; maytansinoid; protein chemistry