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. 2002 Oct;8(5):863-7.
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  1 , Jun Cheng, Gang Wang, Shuang-Shuang Shi, Li Li, Ling-Xia Zhang, Ju-Mei Chen
Affiliations

Preparation of human single chain Fv antibody against hepatitis C virus E2 protein and its identification in immunohistochemistry

Yan-Wei Zhong et al. World J Gastroenterol. 2002 Oct.

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 SfiI/NotI 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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Figures

Figure 1
Figure 1
Restriction map of HCV E2-scFv by Sfi I/Not I digestion. A, B: HCV E2-scFv; C: DNA Marker.
Figure 2
Figure 2
Nucleic acid and deduced amino acid sequences of scFv for HCV E2 protein GenBank accession number for this sequence is AF317001.
Figure 3
Figure 3
Absorbances of HCV-E2-scFv binding to E2 antigen by ELISA. (a). supernant from induced XL1-blue transformed with pCANTAB5E-E2-scFv; (b). posive control; (c). supernant from non-induced XL1-blue transformed with pCANTAB5E-E2-scFv, (d). negtive control.
Figure 4
Figure 4
A. Immunohistochemistry of liver tissue from health person; B. Immunohistochemistry of liver tissue from patients with chronic hepatitis C E2 antigen was detected in the cytoplasm of some liver cells.
See this image and copyright information in PMC

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References

    1. Assy N, Minuk G. A comparison between previous and present histologic assessments of chronic hepatitis C viral infections in humans. World J Gastroenterol. 1999;5:107–110. - PMC - PubMed
    1. Caselmann WH, Serwe M, Lehmann T, Ludwig J, Sproat BS, Engels JW. Design, delivery and efficacy testing of therapeutic nucleic acidsused to inhibit hepatitis C virus gene expression in vitro and in vivo. World J Gastroenterol. 2000;6:626–629. - PMC - PubMed
    1. Cheng JL, Liu BL, Zhang Y, Tong WB, Yan Z, Feng BF. Hepatitis C virus in human B lymphocytes transformed by Epstein-Barr virus in vitro by in situ reverse transcriptase-polymerase chain reaction. World J Gastroenterol. 2001;7:370–375. - PMC - PubMed
    1. Dai YM, Shou ZP, Ni CR, Wang NJ, Zhang SP. Localization of HCV RNA and capsid protein in human hepatocellular carcinoma. World J Gastroenterol. 2000;6:136–137. - PMC - PubMed
    1. Deng ZL, Ma Y, Yuan L, Teng PK. The importance of hepatitis C as a risk factor for hepatocellular carcinoma in Guangxi. World J Gastroenterol. 2000;6(Suppl 3):75.

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