High-Throughput B Cell Epitope Determination by Next-Generation Sequencing
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 po...
| Main Authors: | , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-03-01
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| Series: | Frontiers in Immunology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2022.855772/full |
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| author | Lauren M. Walker Lauren M. Walker Andrea R. Shiakolas Andrea R. Shiakolas Rohit Venkat Zhaojing Ariel Liu Steven Wall Steven Wall Nagarajan Raju Nagarajan Raju Kelsey A. Pilewski Kelsey A. Pilewski Ian Setliff Amyn A. Murji Rebecca Gillespie Nigel A. Makoah Nigel A. Makoah Masaru Kanekiyo Mark Connors Lynn Morris Lynn Morris Lynn Morris Ivelin S. Georgiev Ivelin S. Georgiev Ivelin S. Georgiev Ivelin S. Georgiev Ivelin S. Georgiev Ivelin S. Georgiev |
| author_facet | Lauren M. Walker Lauren M. Walker Andrea R. Shiakolas Andrea R. Shiakolas Rohit Venkat Zhaojing Ariel Liu Steven Wall Steven Wall Nagarajan Raju Nagarajan Raju Kelsey A. Pilewski Kelsey A. Pilewski Ian Setliff Amyn A. Murji Rebecca Gillespie Nigel A. Makoah Nigel A. Makoah Masaru Kanekiyo Mark Connors Lynn Morris Lynn Morris Lynn Morris Ivelin S. Georgiev Ivelin S. Georgiev Ivelin S. Georgiev Ivelin S. Georgiev Ivelin S. Georgiev Ivelin S. Georgiev |
| author_sort | Lauren M. Walker |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-13T07:57:02Z |
| format | Article |
| id | doaj.art-b4cc54924c22433d9477b4b78981ae3b |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-13T07:57:02Z |
| publishDate | 2022-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-b4cc54924c22433d9477b4b78981ae3b2022-12-21T23:54:32ZengFrontiers Media S.A.Frontiers in Immunology1664-32242022-03-011310.3389/fimmu.2022.855772855772High-Throughput B Cell Epitope Determination by Next-Generation SequencingLauren M. Walker0Lauren M. Walker1Andrea R. Shiakolas2Andrea R. Shiakolas3Rohit Venkat4Zhaojing Ariel Liu5Steven Wall6Steven Wall7Nagarajan Raju8Nagarajan Raju9Kelsey A. Pilewski10Kelsey A. Pilewski11Ian Setliff12Amyn A. Murji13Rebecca Gillespie14Nigel A. Makoah15Nigel A. Makoah16Masaru Kanekiyo17Mark Connors18Lynn Morris19Lynn Morris20Lynn Morris21Ivelin S. Georgiev22Ivelin S. Georgiev23Ivelin S. Georgiev24Ivelin S. Georgiev25Ivelin S. Georgiev26Ivelin S. Georgiev27Vanderbilt Vaccine Center, Vanderbilt University Medical Center, Nashville, TN, United StatesDepartment of Pathology, Microbiology and Immunology, Vanderbilt University Medical Center, Nashville, TN, United StatesVanderbilt Vaccine Center, Vanderbilt University Medical Center, Nashville, TN, United StatesDepartment of Pathology, Microbiology and Immunology, Vanderbilt University Medical Center, Nashville, TN, United StatesVanderbilt Vaccine Center, Vanderbilt University Medical Center, Nashville, TN, United StatesVanderbilt Vaccine Center, Vanderbilt University Medical Center, Nashville, TN, United StatesVanderbilt Vaccine Center, Vanderbilt University Medical Center, Nashville, TN, United StatesDepartment of Pathology, Microbiology and Immunology, Vanderbilt University Medical Center, Nashville, TN, United StatesVanderbilt Vaccine Center, Vanderbilt University Medical Center, Nashville, TN, United StatesDepartment of Pathology, Microbiology and Immunology, Vanderbilt University Medical Center, Nashville, TN, United StatesVanderbilt Vaccine Center, Vanderbilt University Medical Center, Nashville, TN, United StatesDepartment of Pathology, Microbiology and Immunology, Vanderbilt University Medical Center, Nashville, TN, United StatesVanderbilt Vaccine Center, Vanderbilt University Medical Center, Nashville, TN, United StatesVanderbilt Vaccine Center, Vanderbilt University Medical Center, Nashville, TN, United StatesVaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health (NIH), Bethesda, MD, United StatesDivision of Virology, Faculty of Health Sciences, University of the Free State, Bloemfontein, South AfricaNational Institute for Communicable Diseases of the National Health Laboratory Service, Johannesburg, South AfricaVaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health (NIH), Bethesda, MD, United StatesNational Institute of Allergy and Infectious Diseases, National Institutes of Health (NIH), Bethesda, MD, United StatesNational Institute for Communicable Diseases of the National Health Laboratory Service, Johannesburg, South AfricaAntibody Immunity Research Unit, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South AfricaCentre for the AIDS Programme of Research in South Africa (CAPRISA), University of KwaZulu-Natal, Durban, South AfricaVanderbilt Vaccine Center, Vanderbilt University Medical Center, Nashville, TN, United StatesDepartment of Pathology, Microbiology and Immunology, Vanderbilt University Medical Center, Nashville, TN, United StatesVanderbilt Institute for Infection, Immunology, and Inflammation, Vanderbilt University Medical Center, Nashville, TN, United States0Department of Electrical Engineering and Computer Science, Vanderbilt University, Nashville, TN, United States1Center for Structural Biology, Vanderbilt University, Nashville, TN, United States2Program in Computational Microbiology and Immunology, Vanderbilt University Medical Center, Nashville, TN, United StatesDevelopment 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.https://www.frontiersin.org/articles/10.3389/fimmu.2022.855772/fullsingle cellepitopemonoclonal antibodyHIVnext generation sequencing (NGS) |
| spellingShingle | Lauren M. Walker Lauren M. Walker Andrea R. Shiakolas Andrea R. Shiakolas Rohit Venkat Zhaojing Ariel Liu Steven Wall Steven Wall Nagarajan Raju Nagarajan Raju Kelsey A. Pilewski Kelsey A. Pilewski Ian Setliff Amyn A. Murji Rebecca Gillespie Nigel A. Makoah Nigel A. Makoah Masaru Kanekiyo Mark Connors Lynn Morris Lynn Morris Lynn Morris Ivelin S. Georgiev Ivelin S. Georgiev Ivelin S. Georgiev Ivelin S. Georgiev Ivelin S. Georgiev Ivelin S. Georgiev High-Throughput B Cell Epitope Determination by Next-Generation Sequencing Frontiers in Immunology single cell epitope monoclonal antibody HIV next generation sequencing (NGS) |
| title | High-Throughput B Cell Epitope Determination by Next-Generation Sequencing |
| title_full | High-Throughput B Cell Epitope Determination by Next-Generation Sequencing |
| title_fullStr | High-Throughput B Cell Epitope Determination by Next-Generation Sequencing |
| title_full_unstemmed | High-Throughput B Cell Epitope Determination by Next-Generation Sequencing |
| title_short | High-Throughput B Cell Epitope Determination by Next-Generation Sequencing |
| title_sort | high throughput b cell epitope determination by next generation sequencing |
| topic | single cell epitope monoclonal antibody HIV next generation sequencing (NGS) |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2022.855772/full |
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