From structure to sequence: Antibody discovery using cryoEM

• Type of Material: Article
• Authors: Aleksander Antanasijevic, Scripps Research Institute; Charles A. Bowman, Scripps Research Institute; Robert N. Kirchdoerfer, University of Wisconsin; Christopher A. Cottrell, Scripps Research Institute; Gabriel Ozorowski, Scripps Research Institute; Amit Upadhyay, Emory University; Kimberly M. Cirelli, La Jolla Institute for Allergy and Immunology; Diane G. Carnathan, Emory University; Chiamaka A. Enemuo, Emory University; Leigh M. Sewall, Scripps Research Institute; Bartrek Nogal, Scripps Research Institute; Fangzhu Zhao, Scripps Research Institute; Bettina Groschel, Scripps Research Institute; William R. Schief, Scripps Research Institute; Devin Sok, Scripps Research Institute; Guido Silvestri, Emory University; Shane Crotty, La Jolla Institute for Allergy and Immunology; Steven Bosinger, Emory University; Andrew B. Ward, Scripps Research Institute
• Language: English
• Date: 2022-01-01
• Publisher: American Association for the Advancement of Science
• Publication Version: Final Published Version
• Copyright Statement: © 2022 The Authors.
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1126/sciadv.abk2039
• Title of Journal or Parent Work: Science Advances
• Volume: 8
• Issue: 3
• Start Page: eabk2039
• End Page: eabk2039
• Grant/Funding Information: Yerkes National Primate Research Center is supported by the base grant P51 OD011132, and the Yerkes Genomics Core by NIH S10 OD026799 awarded to S.E.B.; R.N.K. is supported by NIH/NIAID K99 AI123498. This work was supported by the IAVI Neutralizing Antibody Center through the CAVD.; Center for HIV/AIDS Vaccine Development UM1AI144462 (A.B.W. and D.S.), P01 AI110657 (A.B.W.), and by the Bill & Melinda Gates Foundation and the Collaboration for AIDS Vaccine Discovery (CAVD) OPP1115782/INV-002916 (A.B.W.).; C.A.C. was supported by an NIH F31 Ruth L. Kirschstein Predoctoral Award AI131873 and by the Achievement Rewards for College Scientists Foundation. A.A. is supported by amfAR Mathilde Krim Fellowships in Biomedical Research no. 110182-69-RKVA.; This work was supported by grants from the National Institute of Allergy and Infectious Diseases, Center for HIV/AIDS Vaccine Immunology and Immunogen Discovery UM1AI100663 (A.B.W. and D.S.)
• Keywords: Responses; Multidisciplinary Sciences; Immunoglobin; Nonhuman primates; Science & Technology - Other Topics; Identification; Generation; Science & Technology; HIV; Technologies; Monoclonal antibodies; Visualization
• Research Categories: Health Sciences, Pathology; Health Sciences, Immunology; Biology, Microbiology

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.