[Establishment of anti-HCV NS5 alpha McAb hybridoma cell lines and its application on clinical immunohistochemical diagnosis].
OBJECTIVE
To prepare hybridoma cell lines which secrete anti-HCV recombinant NS5a protein monoclonal antibodies, and to evaluate their usage in investigating the distribution of HCV NS5a antigen in liver tissue.


METHODS
The hybridoma cell lines were raised using the conventional protocols. The immunohistochemistry of paraffin embedded liver tissue was performed to determine the distribution of HCV NS5a antigen as well as NS3 antigen in liver tissue.


RESULTS
Four hybridoma cell lines, which secrete monoclonal antibodies against HCV NS5a protein, were raised and named as 8B2, 6F11, 4C6 and 7D9. They had specific reaction with HCV NS5a protein and had no reaction with HCV core and NS3 proteins. Immunohistochemistry results indicated that 54.8% of anti-HCV positive patient's liver specimens were positive and 70.6% of active cirrhosis positive.


CONCLUSIONS
The monoclonal antibodies we prepared here are specific to the recombinant HCV NS5 alpha protein and they can be used in the clinical immunohistochemistry diagnosis.
Gao
J
J
Hepatology Institute, Beijing Medical University.
Ji
H
H
Lang
Z
Z
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D016174:Hepacivirus / Q000302:isolation & purification*; D006526:Hepatitis C / Q000175:diagnosis*; D018937:Hepatitis C Antibodies / Q000096:biosynthesis*; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000096:biosynthesis; D017361:Viral Nonstructural Proteins / Q000032:analysis / Q000276:immunology*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; D012324:RNA-Dependent RNA Polymerase
D000911:Antibodies, Monoclonal; D018937:Hepatitis C Antibodies; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase