[The preparation of human single chain Fv antibody specifically against hepatitis C virus NS3 antigen and its application in immunohistochemistry].
OBJECTIVE
Immunohistochemistry identification of human single chain Fv antibody (ScFv) against hepatitis C viral NS3 antigen.


METHODS
The recombinant phages were panned by NS3 antigen which was coated on a microtiter plate, after five rounds of biopanning, 66 phage clones were identified specific to NS3 antigen. The affinity and specificity of ScFv were evaluated by ELISA, dot blot hybridization and immunohistochemistry.


RESULTS
The data of ScFv-NS3 DNA digestion and DNA sequencing showed that the ScFv gene is composed of 747 bp. ELISA, dot blot hybridization and immunohistochemistry demonstrated that the human single chain Fv antibody against hepatitis C NS3 antigen has a specific combination character with hepatitis C virus NS3 antigen of different sources and paraffin-embedded patients tissues, it didn't react with liver tissue and HBsAg.


CONCLUSIONS
The application of HCV NS3 specific ScFv in immunohistochemistry was successfully achieved.
Zhong
Y
Y
Gene Therapy Research Centre, Institute of Infectious Diseases, 302 Hospital of PLA, Beijing 100039, China.
Wang
S
S
Zhao
J
J
D002478:Cells, Cultured; D016174:Hepacivirus / Q000276:immunology*; D006801:Humans; D007128:Immunoglobulin Fragments / Q000096:biosynthesis*; D007150:Immunohistochemistry; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000276:immunology*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D007128:Immunoglobulin Fragments; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C055966:immunoglobulin Fv