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


CONCLUSIONS
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.
Zhong
Yan-Wei
YW
Gene Therapy Research Center, Institute of Infectious Diseases, 302 Hospital of PLA, 26 Fengtai Road, Beijing 100039, China. zyw@genetherapy.com.cn
Cheng
Jun
J
Wang
Gang
G
Shi
Shuang-Shuang
SS
Li
Li
L
Zhang
Ling-Xia
LX
Chen
Ju-Mei
JM
D000595:Amino Acid Sequence; D000918:Antibody Specificity; D001435:Bacteriophages; D001483:Base Sequence; D004926:Escherichia coli; D015870:Gene Expression; D015723:Gene Library; D005820:Genetic Testing; D019698:Hepatitis C, Chronic / Q000473:pathology*; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics* / Q000276:immunology; D007150:Immunohistochemistry; D008099:Liver / Q000473:pathology / Q000821:virology; D008969:Molecular Sequence Data; D014759:Viral Envelope Proteins / Q000032:analysis* / Q000276:immunology*
D007128:Immunoglobulin Fragments; D014759:Viral Envelope Proteins; C055966:immunoglobulin Fv; C091015:glycoprotein E2, Hepatitis C virus