title: Broad epitope coverage of a human in vitro antibody library.
abstract: Successful discovery of therapeutic antibodies hinges on the identification of appropriate affinity binders targeting a diversity of molecular epitopes presented by the antigen. Antibody campaigns that yield such broad "epitope coverage" increase the likelihood of identifying candidates with the desired biological functions. Accordingly, epitope binning assays are employed in the early discovery stages to partition antibodies into epitope families or "bins" and prioritize leads for further characterization and optimization. The collaborative program described here, which used hen egg white lysozyme (HEL) as a model antigen, combined 3 key capabilities: 1) access to a diverse panel of antibodies selected from a human in vitro antibody library; 2) application of state-of-the-art high-throughput epitope binning; and 3) analysis and interpretation of the epitope binning data with reference to an exhaustive set of published antibody:HEL co-crystal structures. Binning experiments on a large merged panel of antibodies containing clones from the library and the literature revealed that the inferred epitopes for the library clones overlapped with, and extended beyond, the known structural epitopes. Our analysis revealed that nearly the entire solvent-exposed surface of HEL is antigenic, as has been proposed for protein antigens in general. The data further demonstrated that synthetic antibody repertoires provide as wide epitope coverage as those obtained from animal immunizations. The work highlights molecular insights contributed by increasingly higher-throughput binning methods and their broad utility to guide the discovery of therapeutic antibodies representing a diverse set of functional epitopes.
authors.affiliation: a Adimab LLC , Lebanon , NH , USA.
authors.forename: Arvind
authors.initials: A
authors.lastname: Sivasubramanian
authors.affiliation: a Adimab LLC , Lebanon , NH , USA.
authors.forename: Patricia
authors.initials: P
authors.lastname: Estep
authors.affiliation: a Adimab LLC , Lebanon , NH , USA.
authors.forename: Heather
authors.initials: H
authors.lastname: Lynaugh
authors.affiliation: a Adimab LLC , Lebanon , NH , USA.
authors.forename: Yao
authors.initials: Y
authors.lastname: Yu
authors.affiliation: b Wasatch Microfluidics, Inc. , Salt Lake City , UT , USA.
authors.forename: Adam
authors.initials: A
authors.lastname: Miles
authors.affiliation: b Wasatch Microfluidics, Inc. , Salt Lake City , UT , USA.
authors.forename: Josh
authors.initials: J
authors.lastname: Eckman
authors.affiliation: a Adimab LLC , Lebanon , NH , USA.
authors.forename: Kevin
authors.initials: K
authors.lastname: Schutz
authors.affiliation: a Adimab LLC , Lebanon , NH , USA.
authors.forename: Crystal
authors.initials: C
authors.lastname: Piffath
authors.affiliation: a Adimab LLC , Lebanon , NH , USA.
authors.forename: Nadthakarn
authors.initials: N
authors.lastname: Boland
authors.affiliation: a Adimab LLC , Lebanon , NH , USA.
authors.forename: Rebecca Hurley
authors.initials: RH
authors.lastname: Niles
authors.affiliation: a Adimab LLC , Lebanon , NH , USA.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Durand
authors.affiliation: a Adimab LLC , Lebanon , NH , USA.
authors.forename: Todd
authors.initials: T
authors.lastname: Boland
authors.affiliation: a Adimab LLC , Lebanon , NH , USA.
authors.forename: Maximiliano
authors.initials: M
authors.lastname: Vásquez
authors.affiliation: a Adimab LLC , Lebanon , NH , USA.
authors.forename: Yingda
authors.initials: Y
authors.lastname: Xu
authors.affiliation: c Rinat-Pfizer, Inc. , South San Francisco, CA , USA.
authors.forename: Yasmina
authors.initials: Y
authors.lastname: Abdiche
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology*; D002642:Chick Embryo; D002645:Chickens; D055808:Drug Discovery / Q000379:methods*; D018604:Epitope Mapping / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D009113:Muramidase / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; C421864:hen egg lysozyme; D009113:Muramidase
keywords: Biosensor; epitope; epitope binning; epitope coverage; hen egg white lysozyme; therapeutic monoclonal antibody