Viral Hepatitis
Copyright ©The Author(s) 2002. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Oct 15, 2002; 8(5): 863-867
Published online Oct 15, 2002. doi: 10.3748/wjg.v8.i5.863
Preparation of human single chain Fv antibody against hepatitis C virus E2 protein and its identification in immunohistochemistry
Yan-Wei Zhong, Jun Cheng, Gang Wang, Shuang-Shuang Shi, Li Li, Ling-Xia Zhang, Ju-Mei Chen
Yan-Wei Zhong, Jun Cheng, Gang Wang, Shuang-Shuang Shi, Li Li, Ling-Xia Zhang, Ju-Mei Chen, Gene Therapy Research Center, Institute of Infectious Diseases, 302 Hospital of PLA, 26 Fengtai Road, Beijing 100039, China
Author contributions: All authors contributed equally to the work.
Supported by the National Natural Science Foundation of China, No.39900130
Correspondence to: Yan-Wei Zhong, Gene Therapy Research Center, Institute of Infectious Diseases, 302 Hospital of PLA, 26 Fengtai Road, Beijing 100039, China. zyw@genetherapy.com.cn
Telephone: +86-10-66933392 Fax: +86-10-63801283
Received: January 28, 2002
Revised: March 1, 2002
Accepted: March 5, 2002
Published online: October 15, 2002
Abstract

AIM: To screen human single chain Fv antibody (scFv) against hepatitis C virus E2 antigen and identify its application in immunohistochemistry.

METHODS: The phage antibody library was panned by HCV E2 antigen, which was coated in microtiter plate. After five rounds of biopanning, 56 phage clones were identified specific to HCV E2 antigen. The selected scFv clones were digested by Sfi I/Not I and DNA was sequenced. Then it was subcloned into the vector pCANTAB5E for expression as E-tagged soluble scFv. The liver tissue sections from normal person and patients with chronic hepatitis B and chronic hepatitis C were immunostained with HCV E2 scFv antibody.

RESULTS: The data of scFv-E2 DNA digestion and DNA sequencing showed that the scFv gene is composed of 750 bp. ELISA and immunohistochemistry demonstrated that the human single chain Fv antibody against hepatitis C E2 antigen has a specific binding character with hepatitis virus E2 antigen and paraffin-embedded tissue, but did not react with liver tissues from healthy persons or patients with chronic hepatitis B.

CONCLUSION: We have successfully screened and identified HCV E2 scFv and the scFv could be used in the immunostaining of liver tissue sections from patients with chronic hepatitis C.

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