Sequence-Based Discovery Demonstrates That Fixed Light Chain Human Transgenic Rats Produce a Diverse Repertoire of Antigen-Specific Antibodies.

Autor:	Harris KE; Teneobio, Inc., Menlo Park, CA, United States., Aldred SF; Teneobio, Inc., Menlo Park, CA, United States., Davison LM; Teneobio, Inc., Menlo Park, CA, United States., Ogana HAN; Teneobio, Inc., Menlo Park, CA, United States., Boudreau A; Teneobio, Inc., Menlo Park, CA, United States., Brüggemann M; Teneobio, Inc., Menlo Park, CA, United States., Osborn M; Teneobio, Inc., Menlo Park, CA, United States., Ma B; Teneobio, Inc., Menlo Park, CA, United States., Buelow B; Teneobio, Inc., Menlo Park, CA, United States., Clarke SC; Teneobio, Inc., Menlo Park, CA, United States., Dang KH; Teneobio, Inc., Menlo Park, CA, United States., Iyer S; Teneobio, Inc., Menlo Park, CA, United States., Jorgensen B; Teneobio, Inc., Menlo Park, CA, United States., Pham DT; Teneobio, Inc., Menlo Park, CA, United States., Pratap PP; Teneobio, Inc., Menlo Park, CA, United States., Rangaswamy US; Teneobio, Inc., Menlo Park, CA, United States., Schellenberger U; Teneobio, Inc., Menlo Park, CA, United States., van Schooten WC; Teneobio, Inc., Menlo Park, CA, United States., Ugamraj HS; Teneobio, Inc., Menlo Park, CA, United States., Vafa O; Teneobio, Inc., Menlo Park, CA, United States., Buelow R; Teneobio, Inc., Menlo Park, CA, United States., Trinklein ND; Teneobio, Inc., Menlo Park, CA, United States.
Jazyk:	angličtina
Zdroj:	Frontiers in immunology [Front Immunol] 2018 Apr 24; Vol. 9, pp. 889. Date of Electronic Publication: 2018 Apr 24 (Print Publication: 2018).
DOI:	10.3389/fimmu.2018.00889
Abstrakt:	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:	MEDLINE
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