Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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Successful Application of the Dual-Vector System II in Creating a Reliable Phage-Displayed Combinatorial Fab Library

  • Song, Suk-yoon (Division of Molecular and Medical Biotechnology, College of Bioscience and Biotechnology, Kangwon National University) ;
  • Hur, Byung-ung (Division of Molecular and Medical Biotechnology, College of Bioscience and Biotechnology, Kangwon National University) ;
  • Lee, Kyung-woo (IG Therapy Co., Kangwon National University) ;
  • Choi, Hyo-jung (Division of Molecular and Medical Biotechnology, College of Bioscience and Biotechnology, Kangwon National University) ;
  • Kim, Sung-soo (Department of Pharmacology, School of Medicine, Kangwon National University) ;
  • Kang, Goo (Department of Pathology, School of Medicine, Kangwon National University) ;
  • Cha, Sang-hoon (Division of Molecular and Medical Biotechnology, College of Bioscience and Biotechnology, Kangwon National University)
  • Received : 2008.09.30
  • Accepted : 2008.12.29
  • Published : 2009.03.31
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Abstract

The dual-vector system-II (DVS-II), which allows efficient display of Fab antibodies on phage, has been reported previously, but its practical applicability in a phage-displayed antibody library has not been verified. To resolve this issue, we created two small combinatorial human Fab antibody libraries using the DVS-II, and isolation of target-specific antibodies was attempted. Biopanning of one antibody library, termed DVFAB-1L library, which has a $1.3{\times}10^7$ combinatorial antibody complexity, against fluorescein-BSA resulted in successful isolation of human Fab clones specific for the antigen despite the presence of only a single light chain in the library. By using the unique feature of the DVS-II, an antibody library of a larger size, named DVFAB-131L, which has a $1.5{\times}10^9$ combinatorial antibody complexity, was also generated in a rapid manner by combining $1.3{\times}10^7$ heavy chains and 131 light chains and more diverse anti-fluorescein-BSA Fab antibody clones were successfully obtained. Our results demonstrate that the DVS-II can be applied readily in creating phage-displayed antibody libraries with much less effort, and target-specific antibody clones can be isolated reliably via light chain promiscuity of antibody molecules.

Keywords

Acknowledgement

Supported by : Small and Medium Business Administration, Kangwon National University

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