Get 20M+ Full-Text Papers For Less Than $1.50/day. Start a 14-Day Trial for You or Your Team.

Learn More →

High-resolution analysis of the B cell repertoire before and after polyethylene glycol fusion reveals preferential fusion of rare antigen-specific B cells

High-resolution analysis of the B cell repertoire before and after polyethylene glycol fusion... OBJECTIVE: The hybridoma technology is one of the most important advances inclinical immunology. Little is known about the differences between theantibodies produced during the in vivo immune response and those recovered inhybridoma libraries. Here, we investigate a potential fusion bias inherentto the hybridoma production process.METHODS: Transgenic rats carrying human Ig heavy and light chain loci wereimmunized with measles virus (MV) to generate human mAbs. Usin ghigh-throughput sequencing of IgH mRNA, we compared the IgH repertoire oflymph nodes and the derived hybridoma library using the sequences of theMV-specific hybridoma clones as a reference set with known specificity.RESULTS: We observed that large clonotypes from the lymph nodes were notrepresented in the hybridoma library, but low-frequency B cell populationsbecame highly enriched and most hybridoma clones were derived from these.Our data also showed that identical CDR3s evolved from diverse VDJrecombinations, indicating convergence of different B cells subpopulationstowards expression of antibodies with similar paratopes.CONCLUSION: The efficient generation of mAbs results from a fusion processhighly selective for rare antigen-specific B cells rather than in vivo expandedpopulations. Antibodies of particular interest may therefore be missedduring classical hybridoma production. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Human Antibodies IOS Press

High-resolution analysis of the B cell repertoire before and after polyethylene glycol fusion reveals preferential fusion of rare antigen-specific B cells

Download PDF
15 pages

Loading next page...
 
/lp/ios-press/high-resolution-analysis-of-the-b-cell-repertoire-before-and-after-jS1CiGk0co

References

References for this paper are not available at this time. We will be adding them shortly, thank you for your patience.

Publisher
IOS Press
Copyright
Copyright © 2016 IOS Press and the authors. All rights reserved
ISSN
1093-2607
eISSN
1875-869X
DOI
10.3233/HAB-150288
pmid
27472868
Publisher site
See Article on Publisher Site

Abstract

OBJECTIVE: The hybridoma technology is one of the most important advances inclinical immunology. Little is known about the differences between theantibodies produced during the in vivo immune response and those recovered inhybridoma libraries. Here, we investigate a potential fusion bias inherentto the hybridoma production process.METHODS: Transgenic rats carrying human Ig heavy and light chain loci wereimmunized with measles virus (MV) to generate human mAbs. Usin ghigh-throughput sequencing of IgH mRNA, we compared the IgH repertoire oflymph nodes and the derived hybridoma library using the sequences of theMV-specific hybridoma clones as a reference set with known specificity.RESULTS: We observed that large clonotypes from the lymph nodes were notrepresented in the hybridoma library, but low-frequency B cell populationsbecame highly enriched and most hybridoma clones were derived from these.Our data also showed that identical CDR3s evolved from diverse VDJrecombinations, indicating convergence of different B cells subpopulationstowards expression of antibodies with similar paratopes.CONCLUSION: The efficient generation of mAbs results from a fusion processhighly selective for rare antigen-specific B cells rather than in vivo expandedpopulations. Antibodies of particular interest may therefore be missedduring classical hybridoma production.

Journal

Human Antibodies – IOS Press

Published: Jan 1, 2016

There are no references for this article.