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KMID : 0578320090270030313
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Molecules and Cells
2009 Volume.27 No. 3 p.313 ~ p.319
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Successful Application of the Dual-Vector Sys-tem II in Creating a Reliable Phage-Displayed Combinatorial Fab Library
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Song Suk-Yoon
Hur Byung-Ung
Lee Kyung-woo
Choi Hyo-jung
Kim Sung-Soo
Kang Goo
Cha Sang-hoon
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Abstract
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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 re-solve this issue, we created two small combinatorial hu-man 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 ??107 combinatorial antibody complexity, against fluorescein-BSA resulted in success-ful 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 ??109 combinatorial antibody complexity, was also generated in a rapid manner by combining 1.3 ??107 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.
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KEYWORD
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dual-vector system, Ex12 helper phage, Fab, human antibody, phage display technology
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