2010年5月28日
Enhancement of hypermutation frequency in the chicken B cell line DT40 for efficient diversification of the antibody repertoire.
Biochemical and biophysical research communications
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- 巻
- 396
- 号
- 2
- 開始ページ
- 353
- 終了ページ
- 8
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1016/j.bbrc.2010.04.096
- 出版者・発行元
- ACADEMIC PRESS INC ELSEVIER SCIENCE
Chicken B cell line DT40 continuously accumulates mutations in the immunoglobulin variable region (IgV) gene by gene conversion and point mutation, both of which are mediated by activation-induced cytidine deaminase (AID), thereby producing an antibody (Ab) library that is useful for screening monoclonal Abs (mAbs) in vitro. We previously generated an engineered DT40 line named DT40-SW, whose AID expression can be reversibly switched on or off, and developed an in vitro Ab generation system using DT40-SW cells. To efficiently create an Ab library with sufficient diversity, higher hypermutation frequency is advantageous. To this end, we generated a novel cell line DT40-SWDeltaC, which conditionally expresses a C-terminus-truncated AID mutant lacking the nuclear export signal. The transcription level of the mutant AID gene in DT40-SWDeltaC cells was similar to that of the wild-type gene in DT40-SW cells. However, the protein level of the truncated AID mutant was less than that of the wild type. The mutant protein was enriched in the nuclei of DT40-SWDeltaC cells, although the protein might be highly susceptible to degradation. In DT40-SWDeltaC cells, both gene conversion and point mutation occurred in the IgV gene with over threefold higher frequency than in DT40-SW cells, suggesting that a lower level of the mutant AID protein was sufficient to increase mutation frequency. Thus, DT40-SWDeltaC cells may be useful for constructing Ab libraries for efficient screening of mAbs in vitro.
- リンク情報
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- DOI
- https://doi.org/10.1016/j.bbrc.2010.04.096
- PubMed
- https://www.ncbi.nlm.nih.gov/pubmed/20416279
- Web of Science
- https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000278658000030&DestApp=WOS_CPL
- Scopus
- https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=77952741054&origin=inward
- Scopus Citedby
- https://www.scopus.com/inward/citedby.uri?partnerID=HzOxMe3b&scp=77952741054&origin=inward
- ID情報
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- DOI : 10.1016/j.bbrc.2010.04.096
- ISSN : 0006-291X
- eISSN : 1090-2104
- PubMed ID : 20416279
- SCOPUS ID : 77952741054
- Web of Science ID : WOS:000278658000030