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A Novel Human scFv Library with Non-Combinatorial Synthetic CDR Diversity

Title
A Novel Human scFv Library with Non-Combinatorial Synthetic CDR Diversity
Authors
Bai, XuelianKim, JihyeKang, SeungminKim, WankyuShim, Hyunbo
Ewha Authors
심현보
SCOPUS Author ID
심현보scopus
Issue Date
2015
Journal Title
PLOS ONE
ISSN
1932-6203JCR Link
Citation
vol. 10, no. 10
Publisher
PUBLIC LIBRARY SCIENCE
Indexed
SCIE; SCOPUS WOS
Abstract
The present work describes the construction and validation of a human scFv library with a novel design approach to synthetic complementarity determining region (CDR) diversification. The advantage of synthetic antibody libraries includes the possibility of exerting fine control over factors like framework sequences, amino acid and codon usage, and CDR diversity. However, random combinatorial synthesis of oligonucleotides for CDR sequence diversity also produces many clones with unnatural sequences and/or undesirable modification motifs. To alleviate these issues, we designed and constructed a novel semi-synthetic human scFv library with non-combinatorial, pre-designed CDR diversity and a single native human framework each for heavy, kappa, and lambda chain variable domains. Next-generation sequencing analysis indicated that the library consists of antibody clones with highly nature-like CDR sequences and the occurrence of the post-translational modification motifs is minimized. Multiple unique clones with nanomolar affinity could be isolated from the library against a number of target antigens, validating the library design strategy. The results demonstrate that it is possible to construct a functional antibody library using low, non-combinatorial synthetic CDR diversity, and provides a new strategy for the design of antibody libraries suitable for demanding applications.
DOI
10.1371/journal.pone.0141045
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일반대학원 > 바이오융합과학과 > Journal papers
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