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Analysis of HBV-Specific CD4 T-cell Responses and Identification of HLA-DR-Restricted CD4 T-Cell Epitopes Based on a Peptide Matrix
Published on: October 20, 2021
[Identification of HCV antigen epitopes from a random 15-peptide library by anti-HCV polycloral antibody]
Shen Ke1, Ji-ping Zhu, Xin-yu Wen
1Institute of Basic Medical Science, PLA General Hospital, Beijing 100853, China.
Aim:
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.
Conclusion:
These findings indicate that WPWS motif in the short peptide may mimic the HCV epitope recognized by anti-HCV Abs.

