[Identification of HCV antigen epitopes from a random 15-peptide library by anti-HCV polycloral antibody].
OBJECTIVE
To analyze epitope recognized by anti-HCV antibodies; patients suffered from hepatitis C.


METHODS
Anti-HCV Abs were purified from the patients serum through an affinity chromatography column which was prepared with sepharose 4B coupled with protein A. These Abs were used for biopanning of a phage-displayed random 15-peptide library.


RESULTS
After 3 rounds of biopanning, the ratio of output to input increased to 3.3 x 10(3) and the false positive rate reduced to 0.2%, suggesting that the enrichment was effective. After the third round of biopanning, sixteen clones were selected to conduct binding test to Abs from the patients and normal person's sera. Nine of them were proved to react specifically to the sera from the patients. From the deduced insert sequence in the coat protein VIII, the core sequence of WPWS was found in 8 clones. The positive phage clones could react to different patients' and not react to normal person's sera.


CONCLUSIONS
These findings indicate that WPWS motif in the short peptide may mimic the HCV epitope recognized by anti-HCV Abs.
Ke
Shen
S
Institute of Basic Medical Science, PLA General Hospital, Beijing 100853, China.
Zhu
Ji-ping
JP
Wen
Xin-yu
XY
Zhao
Ping
P
Shi
Ming
M
He
Yong-huai
YH
Yao
Zhi-jian
ZJ
Shen
Bei-fen
BF
D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000276:immunology*; D018604:Epitope Mapping / Q000379:methods; D000939:Epitopes / Q000276:immunology*; D016174:Hepacivirus / Q000276:immunology*; D006526:Hepatitis C / Q000276:immunology; D018937:Hepatitis C Antibodies / Q000276:immunology* / Q000302:isolation & purification; D018936:Hepatitis C Antigens / Q000276:immunology*; D006801:Humans; D008969:Molecular Sequence Data; D019151:Peptide Library
D000939:Epitopes; D018937:Hepatitis C Antibodies; D018936:Hepatitis C Antigens; D019151:Peptide Library