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Successful application of the dual-vector system II in creating a reliable phage-displayed combinatorial Fab library.

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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 x 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 x 10(9) combinatorial antibody complexity, was also generated in a rapid manner by combining 1.3 x 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 molecule.

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      Author and article information

      Affiliations
      [1 ] Division of Molecular and Medical Biotechnology, College of Bioscience and Biotechnology, Kangwon National University, Chuncheon 200-701, Korea.
      Journal
      Mol. Cells
      Molecules and cells
      Springer Nature America, Inc
      0219-1032
      1016-8478
      Mar 31 2009
      : 27
      : 3
      19326078
      10.1007/s10059-009-0040-0

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