Screening for PreS specific binding ligands with a phage displayed peptides library.
OBJECTIVE
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+/-10(8) 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.


CONCLUSIONS
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.
Deng
Qiang
Q
Institute of Biochemistry and Cell Biology, 320 Yue-Yang Road, Shanghai 200031, China.
Zhuang
Ming
M
Kong
Yu-Ying
YY
Xie
You-Hua
YH
Wang
Yuan
Y
D000595:Amino Acid Sequence; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular; D045744:Cell Line, Tumor; D017931:DNA Primers; D006514:Hepatitis B Surface Antigens / Q000235:genetics / Q000502:physiology*; D006515:Hepatitis B virus / Q000235:genetics / Q000276:immunology*; D006801:Humans; D008113:Liver Neoplasms; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry; D019151:Peptide Library; D011498:Protein Precursors / Q000235:genetics / Q000502:physiology*; D015183:Restriction Mapping; D014759:Viral Envelope Proteins / Q000502:physiology
D017931:DNA Primers; D006514:Hepatitis B Surface Antigens; D009842:Oligopeptides; D019151:Peptide Library; D011498:Protein Precursors; D014759:Viral Envelope Proteins; C050807:presurface protein 2, hepatitis B surface antigen