title: Successful application of the dual-vector system II in creating a reliable phage-displayed combinatorial Fab library.
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.
authors.affiliation: Division of Molecular and Medical Biotechnology, College of Bioscience and Biotechnology, Kangwon National University, Chuncheon 200-701, Korea.
authors.forename: Suk-yoon
authors.initials: SY
authors.lastname: Song
authors.forename: Byung-ung
authors.initials: BU
authors.lastname: Hur
authors.forename: Kyung-woo
authors.initials: KW
authors.lastname: Lee
authors.forename: Hyo-jung
authors.initials: HJ
authors.lastname: Choi
authors.forename: Sung-soo
authors.initials: SS
authors.lastname: Kim
authors.forename: Goo
authors.initials: G
authors.lastname: Kang
authors.forename: Sang-hoon
authors.initials: SH
authors.lastname: Cha