Production and characterization of monoclonal antibody specific for NS3 helicase of hepatitis C virus.
Hepatitis C virus (HCV) infection is the major etiological agent of chronic hepatitis, which leads to liver cirrhosis and hepatocellular carcinomas. HCV NS3 helicase is a promising target of anti-virus therapy. In this report, we discuss a strategy to generate monoclonal antibodies (MAbs) of the HCV NS3 helicase, and investigate its potential characteristic. Our results showed the production of MAbs against NS3 helicase, which could specifically recognize the native NS3 helicase in transiently transfected cells in the immunofluorescence experiment. The resultant MAbs were used as the first antibody in Western blot analyses, and observed the specific band that defines the NS3 helicase. Likewise, one MAb could inhibit the NS3 helicase enzymatic activity distinctly in the NS3 helicase-mediated DNA-unwinding assay. To conclude, these antibodies may be useful to generate specific diagnostic tools for HCV infection and may also be developed for potential therapeutics.
Yang
Jing
J
The State Key Discipline and Department of Microbiology Fourth Medical Military University, 17th Changlexi Road, Xi'an 710032, China.
Lei
Ying-Feng
YF
Yin
Wen
W
Wei
San-Hua
SH
An
Qun-Xing
QX
Lv
Xin
X
Hu
Xing-Bin
XB
Xu
Zhi-Kai
ZK
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D015153:Blotting, Western; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D019084:Fluorescent Antibody Technique, Indirect; D016174:Hepacivirus / Q000378:metabolism*; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D007107:Immune System; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence / Q000379:methods; D010957:Plasmids / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000737:chemistry*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D000911:Antibodies, Monoclonal; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases