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
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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