摘要
AIM: To screen human single chain Fv antibody (scFv)against hepatitis C virus E2 antigen and identify its applicationin immunohistochemistry.METHODS: The phage antibody library was panned by HCVE2 antigen, which was coated in microtiter plate. After fiverounds of biopanning,56 phage clones were identified specificto HCV E2 antigen. The selected scFv clones were digestedby SfiI/NotI and DNA was sequenced. Then it was subclonedinto the vector pCANTABSE for expression as E-taggedsoluble scFv. The liver tissue sections from normal personand patients with chronic hepatitis B and chronic hepatitis Cwere immunostained with HCV E2 scFv antibody.RESULTS: The data of scFv-E2 DNA digestion and DNAsequencing showed that the scFv gene is composed of 750bp. ELISA and immunohistochemistry demonstrated that thehuman single chain Fy antibody against hepatitis C E2 antigenhas a specific binding character with hepatitis virus E2 antigenand paraffin-embedded tissue, but did not react with liver tissuesfrom healthy persons or patients with chronic hepatitis B.CONCLUSION: We have successfully screened andidentified HCV E2 scFv and the scFv could be used in theimmunostaining of liver tissue sections from patients withchronic hepatitis C.
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 SflI/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.
基金
the National Natural Science Foundation of China,No.39900130
