Production and characterization of monoclonal antibodies to a grouper iridovirus.
A panel of six monoclonal antibodies (mAbs) against a grouper iridovirus (SGIV) were produced by immunization of Balb/c mice with purified virus preparations. Isotyping test revealed all the mAbs were IgG1. None of the mAbs possessed the ability to neutralize SGIV in cell cultures. Western blot showed that 4 mAbs reacted with 2 SGIV proteins at molecular mass of approximately 100 and 117 kDa in gradient-purified virus. Immunofluorescent studies showed that the two specific viral proteins VP100 and VP117 were localized within virus assembly sites in the cytoplasm of SGIV-infected grouper cells (GP). Fractionations of the iridovirus in a 20-60% sucrose gradient were detected successfully by all the six mAbs using immunodot blot. An antigen-capture enzyme-linked immunosorbent assay (ELISA) system, based on the use of mAb 7E11 for capture and a rabbit polyclonal antibody to SGIV for detection was developed. SGIV antigen was detected in gradient-purified virus and virus-infected grouper blood. These novel mAbs will facilitate the development of more specific and standardized diagnostic techniques for marine fish iridovirus.
Shi
Chengyin
C
Tropical Marine Science Institute, The National University of Singapore, 14 Kent Ridge Road, 119223, Singapore, Singapore.
Wei
Qi
Q
Gin
Karina Yew Hoong
KY
Lam
Toong Jin
TJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology*; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000276:immunology; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000032:analysis / Q000276:immunology; D002460:Cell Line; D004266:DNA Virus Infections / Q000175:diagnosis / Q000662:veterinary*; D004797:Enzyme-Linked Immunosorbent Assay; D005393:Fish Diseases / Q000175:diagnosis* / Q000821:virology; D005455:Fluorescent Antibody Technique; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000276:immunology; D017994:Iridovirus / Q000276:immunology*; D051379:Mice; D010473:Perciformes
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D007074:Immunoglobulin G