Sequence-Based Discovery Demonstrates That Fixed Light Chain Human Transgenic Rats Produce a Diverse Repertoire of Antigen-Specific Antibodies
Autor: | Laura Davison, Starlynn Clarke, Michael J. Osborn, Andrew Boudreau, Kevin Dang, Katherine E. Harris, Biao Ma, Ute Schellenberger, Heather Ogana, Omid Vafa, Nathan D. Trinklein, Roland Buelow, Marianne Brüggemann, Wim Van Schooten, Payal Pratap, Duy Pham, Brett Jorgensen, Benjamin Buelow, Harshad Ugamraj, Shelley Force Aldred, Udaya Rangaswamy, Suhasini Iyer |
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Rok vydání: | 2018 |
Předmět: |
lcsh:Immunologic diseases. Allergy
0301 basic medicine rearranged light chain medicine.drug_class Genes Immunoglobulin Heavy Chain Immunology Somatic hypermutation Context (language use) Computational biology Monoclonal antibody Immunoglobulin light chain monoclonal human antibodies Rats Sprague-Dawley Immunoglobulin kappa-Chains deep sequencing 03 medical and health sciences Antigen Antibody Repertoire transgenic rodent Drug Discovery antibody repertoire medicine Animals Immunology and Allergy Antigens Original Research B-Lymphocytes biology Repertoire Antibodies Monoclonal High-Throughput Nucleotide Sequencing Germinal Center Rats somatic hypermutation 030104 developmental biology humanized rodent Models Animal biology.protein Genes Immunoglobulin Light Chain Rats Transgenic Antibody Immunoglobulin Heavy Chains lcsh:RC581-607 |
Zdroj: | Frontiers in Immunology, Vol 9 (2018) Frontiers in Immunology |
ISSN: | 1664-3224 |
Popis: | We created a novel transgenic rat that expresses human antibodies comprising a diverse repertoire of heavy chains with a single common rearranged kappa light chain (IgKV3-15-JK1). This fixed light chain animal, called OmniFlic, presents a unique system for human therapeutic antibody discovery and a model to study heavy chain repertoire diversity in the context of a constant light chain. The purpose of this study was to analyze heavy chain variable gene usage, clonotype diversity, and to describe the sequence characteristics of antigen-specific monoclonal antibodies (mAbs) isolated from immunized OmniFlic animals. Using next-generation sequencing antibody repertoire analysis, we measured heavy chain variable gene usage and the diversity of clonotypes present in the lymph node germinal centers of 75 OmniFlic rats immunized with 9 different protein antigens. Furthermore, we expressed 2,560 unique heavy chain sequences sampled from a diverse set of clonotypes as fixed light chain antibody proteins and measured their binding to antigen by ELISA. Finally, we measured patterns and overall levels of somatic hypermutation in the full B-cell repertoire and in the 2,560 mAbs tested for binding. The results demonstrate that OmniFlic animals produce an abundance of antigen-specific antibodies with heavy chain clonotype diversity that is similar to what has been described with unrestricted light chain use in mammals. In addition, we show that sequence-based discovery is a highly effective and efficient way to identify a large number of diverse monoclonal antibodies to a protein target of interest. |
Databáze: | OpenAIRE |
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