Expression and detection of ScFvB9 and its mutant in recombinant phage antibody system

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Title
Expression and detection of ScFvB9 and its mutant in recombinant phage antibody system
Author(s)
Eun Ju Yang; Ju Won Kwak; Hae Choon Chang
Bibliographic Citation
Hybridoma and Hybridomics, vol. 20, no. 6, pp. 369-375
Publication Year
2001
Abstract
Recombinant single-chain antibody (ScFvB9) and its mutant (ScFvB9-6) were generated by using a polymerase chain reaction (PCR) from the Fab fragment of the murine monoclonal antibody (MAb) B9, MabB9 (γ2b, κ), which is specific for human plasma apolipoprotein (apo) B-100 of low density lipopreotein (LDL). In the recombinant phage antibody system (RPAS), the constructed ScFvB9 and ScFvB9-6 antibody genes were cloned into the pCANTAB5E phagemid vector and expressed in E. coli. The active forms of single-chain antibodies (ScFvB9 and ScFvB9-6) were produced as phage-displayed recombinant antibodies or soluble antibody forms in E. coli. The activities of ScFvB9 and ScFvB9-6 were confirmed by enzyme-linked immunosorbent assay (ELISA) and Western blotting analysis; the generated mutant ScFvB9-6 showed slightly higher antigen binding activity than native ScFvB9 as a soluble antibody in this RPAS.
ISSN
1536-8599
Publisher
Mary Ann Liebert
DOI
http://dx.doi.org/10.1089/15368590152740770
Type
Article
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1. Journal Articles > Journal Articles
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