View : 63 Download: 0

Construction of a large synthetic human scFv library with six diversified CDRs and high functional diversity

Title
Construction of a large synthetic human scFv library with six diversified CDRs and high functional diversity
Authors
Yang H.Y.Kang K.J.Chung J.E.Shim H.
Ewha Authors
심현보
SCOPUS Author ID
심현보scopus
Issue Date
2009
Journal Title
Molecules and Cells
ISSN
1016-8478JCR Link
Citation
vol. 27, no. 2, pp. 225 - 235
Indexed
SCI; SCIE; SCOPUS; KCI WOS scopus
Abstract
Antibody phage display provides a powerful and efficient tool for the discovery and development of monoclonal antibodies for therapeutic and other applications. Antibody clones from synthetic libraries with optimized design features have several distinct advantages that include high stability, high levels of expression, and ease of downstream optimization and engineering. In this study, a fully synthetic human scFv library with six diversified CDRs was constructed by polymerase chain reaction assembly of overlapping oligonucleotides. In order to maximize the functional diversity of the library, a β-lactamase selection strategy was employed in which the assembled scFv gene repertoire was fused to the 5′-end of the β-lactamase gene, and in-frame scFv clones were enriched by carbenicillin selection. A final library with an estimated total diversity of 7.6 × 109, greater than 70% functional diversity, and diversification of all six CDRs was obtained after insertion of fully randomized CDR-H3 sequences into this proofread repertoire. The performance of the library was validated using a number of target antigens, against which multiple unique scFv sequences with dissociation constants in the nanomolar range were isolated. © 2009 The Korean Society for Molecular and Cellular Biology and Springer Netherlands.
DOI
10.1007/s10059-009-0028-9
Appears in Collections:
일반대학원 > 바이오융합과학과 > Journal papers
Files in This Item:
There are no files associated with this item.
Export
RIS (EndNote)
XLS (Excel)
XML


qrcode

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

BROWSE