Viral Hepatitis
Copyright ©The Author(s) 2005. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jul 14, 2005; 11(26): 4018-4023
Published online Jul 14, 2005. doi: 10.3748/wjg.v11.i26.4018
Screening for PreS specific binding ligands with a phage displayed peptides library
Qiang Deng, Ming Zhuang, Yu-Ying Kong, You-Hua Xie, Yuan Wang
Qiang Deng, Ming Zhuang, Yu-Ying Kong, You-Hua Xie, Yuan Wang, State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes of Life Science, Chinese Academy of Sciences, Shanghai 200031, China
Qiang Deng, You-Hua Xie, Yuan Wang, Sino-France Center for Life Science and Genome Research
Author contributions: All authors contributed equally to the work.
Supported by Basic Research Program from Ministry of Science and Technology of China, No. G1999054105, and special funds for Sino-France Center for Life Science and Genome Research from Chinese Academy of Sciences and Pasteur Institute in France
Correspondence to: Yuan Wang, Institute of Biochemistry and Cell Biology, 320 Yue-Yang Road, Shanghai 200031, China. wangy@sibs.ac.cn
Telephone: +86-21-54921103 Fax: +86-21-54921011
Received: May 29, 2004
Revised: July 11, 2004
Accepted: July 11, 2004
Published online: July 14, 2005
Abstract

AIM: To construct a random peptide phage display library and search for peptides that specifically bind to the PreS region of hepatitis B virus (HBV).

METHODS: A phage display vector, pFuse8, based on the gene 8 product (pVIII) of M13 phage was made and used to construct a random peptide library. E.coli derived thioredoxin-PreS was purified with Thio-bond beads, and exploited as the bait protein for library screening. Five rounds of bio-panning were performed. The PreS-binding specificities of enriched phages were characterized with phage ELISA assay.

RESULTS: A phage display vector was successfully constructed as demonstrated to present a pVIII fused HBV PreS1 epitope on the phage surface with a high efficiency. A cysteine confined random peptide library was constructed containing independent clones exceeding 5±108 clone forming unit (CFU). A pool of phages showing a PreS-binding specificity was obtained after the screening against thio-PreS with an enrichment of approximately 400 times. Five phages with high PreS-binding specificities were selected and characterized. Sequences of the peptides displayed on these phages were determined.

CONCLUSION: A phage library has been constructed, with random peptides displaying as pVIII-fusion proteins. Specific PreS-binding peptides have been obtained, which may be useful for developing antivirals against HBV infection.

Keywords: Hepatitis B virus; Phage; Peptide