From structure to sequence: Antibody discovery using cryoEM.

Antanasijevic, Aleksandar; Bowman, Charles A; Kirchdoerfer, Robert N; Cottrell, Christopher A; Ozorowski, Gabriel; Upadhyay, Amit A; Cirelli, Kimberly M; Carnathan, Diane G et al. · Sci Adv · 2022

basic_science · Level V

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Abstract

One of the rate-limiting steps in analyzing immune responses to vaccines or infections is the isolation and characterization of monoclonal antibodies. Here, we present a hybrid structural and bioinformatic approach to directly assign the heavy and light chains, identify complementarity-determining regions, and discover sequences from cryoEM density maps of serum-derived polyclonal antibodies bound to an antigen. When combined with next-generation sequencing of immune repertoires, we were able to specifically identify clonal family members, synthesize the monoclonal antibodies, and confirm that they interact with the antigen in a manner equivalent to the corresponding polyclonal antibodies. This structure-based approach for identification of monoclonal antibodies from polyclonal sera opens new avenues for analysis of immune responses and iterative vaccine design.