Production and characterization of a monoclonal antibody specific for NS3 protease and the ATPase region of Dengue-2 virus.
Dengue is considered a reemerging disease of worldwide distribution. The Dengue virus non-structural protein 3 (NS3) is known to possess ATPase, helicase, and protease activities that are a constitutive part of the replication complex of Dengue virus. In this report, we discuss the cloning, expressing, and purifying of the Dengue-2 NS3 protein, to immunize mice and then generate monoclonal antibodies (MAbs). Our results show the production of MAbs specific to NS3 protein of Dengue-2 virus, which by immunofluorescence recognize the native protein in experimentally infected endothelial cells (HMEC). Likewise, C6/36-infected lisates were used in Western blots, and observed the specific characteristic band that defines the NS3 protein. We conclude that these antibodies may be a useful tool, not only to study the replicative process of Dengue virus, but also to generate specific diagnostic tools for Dengue infection.
García-Cordero
J
J
Departamento de Biomedicina Molecular, Cinvestav-IPN, Mexico City, Mexico.
Ramirez
H Romero
HR
Vazquez-Ochoa
M
M
Gutierrez-Castañeda
B
B
Santos-Argumedo
L
L
Villegas-Sepúlveda
N
N
Cedillo-Barrón
L
L
D000251:Adenosine Triphosphatases / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D015153:Blotting, Western; D003001:Cloning, Molecular; D003716:Dengue Virus / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D042783:Endothelial Cells / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D019084:Fluorescent Antibody Technique, Indirect; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000378:metabolism; D012697:Serine Endopeptidases / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D000085242:Viral Proteases
D000251:Adenosine Triphosphatases; D000911:Antibodies, Monoclonal; D005982:Glutathione Transferase; D011993:Recombinant Fusion Proteins; D012697:Serine Endopeptidases; C504449:NS3 protease, dengue virus; D000085242:Viral Proteases