High-Throughput B Cell Epitope Determination by Next-Generation Sequencing

Frontiers in Immunology

Ivelin S. Georgiev³  Lynn Morris⁵  Steven Wall⁶  Nagarajan Raju⁶  Lauren M. Walker⁶  Andrea R. Shiakolas⁶  Kelsey A. Pilewski⁶  Nigel A. Makoah⁸  Mark Connors⁹  Masaru Kanekiyo^1,10  Rebecca Gillespie^1,10  Rohit Venkat^1,12  Amyn A. Murji^1,12  Zhaojing Ariel Liu^1,12  Ian Setliff^1,12 

[1] 0Department of Electrical Engineering and Computer Science, Vanderbilt University, Nashville, TN, United States;1Center for Structural Biology, Vanderbilt University, Nashville, TN, United States;2Program in Computational Microbiology and Immunology, Vanderbilt University Medical Center, Nashville, TN, United States;Antibody Immunity Research Unit, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa;Centre for the AIDS Programme of Research in South Africa (CAPRISA), University of KwaZulu-Natal, Durban, South Africa;Department of Pathology, Microbiology and Immunology, Vanderbilt University Medical Center, Nashville, TN, United States;Division of Virology, Faculty of Health Sciences, University of the Free State, Bloemfontein, South Africa;National Institute for Communicable Diseases of the National Health Laboratory Service, Johannesburg, South Africa;National Institute of Allergy and Infectious Diseases, National Institutes of Health (NIH), Bethesda, MD, United States;Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health (NIH), Bethesda, MD, United States;Vanderbilt Institute for Infection, Immunology, and Inflammation, Vanderbilt University Medical Center, Nashville, TN, United States;Vanderbilt Vaccine Center, Vanderbilt University Medical Center, Nashville, TN, United States;

关键词: single cell;    epitope;    monoclonal antibody;    HIV;    next generation sequencing (NGS);   

DOI  :  10.3389/fimmu.2022.855772

来源: DOAJ

【 摘 要 】

Development of novel technologies for the discovery of human monoclonal antibodies has proven invaluable in the fight against infectious diseases. Among the diverse antibody repertoires elicited by infection or vaccination, often only rare antibodies targeting specific epitopes of interest are of potential therapeutic value. Current antibody discovery efforts are capable of identifying B cells specific for a given antigen; however, epitope specificity information is usually only obtained after subsequent monoclonal antibody production and characterization. Here we describe LIBRA-seq with epitope mapping, a next-generation sequencing technology that enables residue-level epitope determination for thousands of single B cells simultaneously. By utilizing an antigen panel of point mutants within the HIV-1 Env glycoprotein, we identified and confirmed antibodies targeting multiple sites of vulnerability on Env, including the CD4-binding site and the V3-glycan site. LIBRA-seq with epitope mapping is an efficient tool for high-throughput identification of antibodies against epitopes of interest on a given antigen target.

【 授权许可】

Unknown