10.3389/fimmu.2018.00889.s001 Katherine E. Harris Katherine E. Harris Shelley Force Aldred Shelley Force Aldred Laura M. Davison Laura M. Davison Heather Anne N. Ogana Heather Anne N. Ogana Andrew Boudreau Andrew Boudreau Marianne Brüggemann Marianne Brüggemann Michael Osborn Michael Osborn Biao Ma Biao Ma Benjamin Buelow Benjamin Buelow Starlynn C. Clarke Starlynn C. Clarke Kevin H. Dang Kevin H. Dang Suhasini Iyer Suhasini Iyer Brett Jorgensen Brett Jorgensen Duy T. Pham Duy T. Pham Payal P. Pratap Payal P. Pratap Udaya S. Rangaswamy Udaya S. Rangaswamy Ute Schellenberger Ute Schellenberger Wim C. van Schooten Wim C. van Schooten Harshad S. Ugamraj Harshad S. Ugamraj Omid Vafa Omid Vafa Roland Buelow Roland Buelow Nathan D. Trinklein Nathan D. Trinklein Image_1_Sequence-Based Discovery Demonstrates That Fixed Light Chain Human Transgenic Rats Produce a Diverse Repertoire of Antigen-Specific Antibodies.jpeg Frontiers 2018 monoclonal human antibodies transgenic rodent rearranged light chain somatic hypermutation deep sequencing antibody repertoire humanized rodent 2018-04-24 04:21:56 Figure https://frontiersin.figshare.com/articles/figure/Image_1_Sequence-Based_Discovery_Demonstrates_That_Fixed_Light_Chain_Human_Transgenic_Rats_Produce_a_Diverse_Repertoire_of_Antigen-Specific_Antibodies_jpeg/6174752 <p>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.</p>