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Nature Biotechnology  23, 731 - 735 (2005)
Published online: 29 May 2005; | doi:10.1038/nbt1092

Rapid generation of specific antibodies by enhanced homologous recombination

Hidetaka Seo1, 2, Mieko Masuoka1, 2, Hiromu Murofushi3, Shunichi Takeda4, Takehiko Shibata5 & Kunihiro Ohta1

1  Genetic Dynamics Research Unit, RIKEN Discovery Research Institute, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.

2  REDS Group (Saitama Prefecture Collaboration of Regional Entities for the Advancement of Technological Excellence, JST), Saitama Small Enterprise Promotion Corporation, 3-12-18 Kamiaoki, Kawaguchi, Saitama 333-0844, Japan.

3  Department of Biology, Faculty of Science, Yamaguchi University, 1677-1 Ohaza Yoshida, Yamaguchi-shi, Yamaguchi 753-8512, Japan.

4  Radiation Genetics, Faculty of Medicine, Kyoto University, Yoshida Konoe, Sakyo-ku, Kyoto 606-8315, Japan.

5  Laboratory of Cellular & Molecular Biology, RIKEN Discovery Research Institute, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.

Correspondence should be addressed to Kunihiro Ohta kohta@riken.jp
In the chicken immune system, gene conversion, a type of homologous recombination, primarily contributes to diversification of the immunoglobulin gene1, 2. Here, we report on the rapid generation of specific monoclonal antibodies using the chicken DT40 B-cell line undergoing gene conversion. We discovered that the gene conversion frequency at the immunoglobulin locus is increased by treating DT40 cells with a histone deacetylase inhibitor, trichostatin A (TSA)3, thereby generating diversity at the immunoglobulin locus in the majority of treated cells. This indicates that TSA treatment accelerates the autonomous diversification of surface IgMs on DT40 cells. We took advantage of this effect to select DT40 cells producing specific antibodies with antigen-conjugated magnetic beads. This autonomously diversifying library (ADLib) selection system enables the quick establishment (approx1 week from a diversifying library) of various clones producing monoclonal IgMs with enough specificity and affinity for immunological assays, and is applicable to various biotechnologies including rational protein design.


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Nature Biotechnology
ISSN: 1087-0156
EISSN: 1546-1696
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