Production of a monoclonal antibody against non-structural protein 3 of dengue-2 virus by intrasplenic injection.
Dengue fever and dengue hemorrhagic fever/dengue shock syndrome are highly infectious diseases caused by dengue virus (DV). DV non-structural protein 3 (NS3) is known to possess ATPase, helicase, and protease activity that is a constitutive part of the replication complex of DV. In this study, we discuss the cloning, expression, and purification of the DV-2 NS3 protein to immunize mice by intrasplenic injection and then to generate a monoclonal antibody (MAb). One MAb, named 4F5, was obtained and it was specific to NS3 of DV-2. Immunofluorescence show that 4F5 recognizes the native protein in infected ECV304 cells. Likewise, C6/36-infected lysates were used in Western blot analysis, and we observed the specific characteristic band that defines NS3. We conclude that MAb 4F5 may be a useful tool, not only to study the replicative process of DV, but also to generate specific diagnostic tools for DV infection.
Chen
Zongtao
Z
Department of Microbiology, Third Military Medical University, Chongqing, P.R. China.
Tian
Yanping
Y
Liu
Limei
L
An
Jing
J
D000330:Aedes; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D003715:Dengue / Q000276:immunology* / Q000821:virology; D003716:Dengue Virus / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods; D051379:Mice; D008807:Mice, Inbred BALB C; D020365:RNA Helicases / Q000737:chemistry / Q000378:metabolism; D012697:Serine Endopeptidases / Q000737:chemistry / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000378:metabolism; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D020365:RNA Helicases; D012697:Serine Endopeptidases; D017361:Viral Nonstructural Proteins; C060028:NS3 protein, flavivirus; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases