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A phage-displayed single chain variable fragment that interacts with hepatitis B core antigen: Library construction, selection and diagnosis

Tan, Geok Hun, Yusoff, Khatijah, Seow, Heng Fong and Tan, Wen Siang (2007) A phage-displayed single chain variable fragment that interacts with hepatitis B core antigen: Library construction, selection and diagnosis. Journal of Clinical Virology, 38 (1). pp. 49-56. ISSN 1386-6532

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Abstract

Background - Phage display is an alternative method for constructing and selecting antibodies with desired specificity towards an antigen. Objectives - To construct a library of single chain variable fragment (ScFv) towards hepatitis B core antigen (HBcAg). To isolate a ScFv phage clone that interacts with HBcAg and to develop a phage-ELISA for detecting the antigen. Study design - Mice were inoculated with HBcAg and RNA was extracted from their spleen cells. The genes encoding heavy (VH) and light (VL) chains were amplified, linked via PCR and cloned into a phagemid vector. Phage particles displaying ScFv were panned against HBcAg and a selected clone was characterized and employed as a diagnostic reagent for detecting HBcAg in serum samples. Results - A phage clone that interacts with HBcAg was selected from the antibody library. The binding of the phage to HBcAg was inhibited by a cyclic peptide bearing the WSFFSNI sequence. A phage-ELISA was established using the recombinant phage and as low as 10 ng of HBcAg can be detected by the assay. Conclusion - The ScFv displayed on the surface of filamentous phage is an alternative choice for diagnosis of HBcAg in serum samples.

Item Type: Article
Uncontrolled Keywords: ResPubID16882, phage display, combinatorial antibody libraries, hepatitis B core antigen, diagnosis, phage-ELISA
Subjects: FOR Classification > 1112 Oncology and Carcinogenesis
Faculty/School/Research Centre/Department > School of Biomedical and Health Sciences
FOR Classification > 0304 Medicinal and Bimolecular Chemistry
SEO Classification > 970111 Expanding Knowledge in the Medical and Health Sciences
Related URLs:
Depositing User: VUIR
Date Deposited: 11 May 2012 01:34
Last Modified: 16 Jan 2015 01:05
URI: http://vuir.vu.edu.au/id/eprint/3417
DOI: https://doi.org/10.1016/j.jcv.2006.09.010
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Citations in Scopus: 5 - View on Scopus

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