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>