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Front. Immunol. | doi: 10.3389/fimmu.2018.00889

Sequence-based discovery demonstrates that fixed light chain human transgenic rats produce a diverse repertoire of antigen-specific antibodies

 Katherine E. Harris1, Shelley F. Aldred1,  Laura M. Davison1, Heather Anne N. Ogana1,  Andrew Boudreau1, Marianne Bruggemann1,  Michael Osborn1,  Biao Ma1, Benjamin Buelow1, Starlynn C. Clarke1, Kevin H. Dang1, Suhasini Iyer1, Brett Jorgensen1, Duy T. Pham1, Payal P. Pratap1, Udaya S. Rangaswamy1,  Ute Schellenberger1, Wim C. van Schooten1, Harshad S. Ugamraj1, Omid Vafa1, Roland Buelow1 and  Nathan Trinklein1*
  • 1Teneobio, Inc., United States

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 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 2560 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. Lastly, we measured patterns and overall levels of somatic hypermutation in the full B-cell repertoire and in the 2560 monoclonal antibodies 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.

Keywords: monoclonal human antibodies, Transgenic rodent, somatic hypermutation, Deep sequencing, Antibody repertoire, humanized rodent, fixed light chain, Antibody discovery, Next generation sequencing (NGS)

Received: 29 Nov 2017; Accepted: 10 Apr 2018.

Edited by:

Bill Harriman, Ligand Pharmaceuticals (United States), United States

Reviewed by:

Alain Lamarre, Institut national de la recherche scientifique (INRS), Canada
Sai T. Reddy, ETH Zürich, Switzerland
Yariv Wine, Tel Aviv University, Israel  

Copyright: © 2018 Harris, Aldred, Davison, Ogana, Boudreau, Bruggemann, Osborn, Ma, Buelow, Clarke, Dang, Iyer, Jorgensen, Pham, Pratap, Rangaswamy, Schellenberger, van Schooten, Ugamraj, Vafa, Buelow and Trinklein. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Dr. Nathan Trinklein, Teneobio, Inc., Menlo Park, United States,