Characterization and application of monoclonal antibodies against turbot (Scophthalmus maximus) Rhabdovirus.
Five monoclonal antibodies (mAbs), 1G8, 1H9, 2D2, 2D3, and 2F5, against Scophthalmus maximus rhabdovirus (SMRV) were prepared. Characterization of the mAbs included indirect enzyme-linked immunosorbent assay, isotyping, viral inhibition assay, immunofluorescence staining of virus-infected cell cultures, and Western blot analysis. Isotyping revealed that 1G8 and 1H9 were of the IgG2b subclass and that the other three were IgM. 2D2, 2D3, and 2F5 partially inhibited SMRV infection in epithelioma papulosum cyprinid (EPC) cell culture. Western blotting showed that all five mAbs could react with two SMRV proteins with molecular masses of approximately 30 kDa (P) and 26 kDa (M). These two proteins were localized within the cytoplasm of SMRV-infected EPC cells by immunofluorescence assay. Also, progressive foci of viral replication in cell cultures were monitored from 6 to 24 h, using mAb 2D3 as the primary antibody. A flow cytometry procedure was used to detect and quantify SMRV-infected (0.01 PFU/cell) EPC cells with mAb 2D3, and 10.8% of cells could be distinguished as infected 36 h postinfection. Moreover, mAb 2D3 was successfully applied for the detection of viral antigen in cryosections from flounder tissues by immunohistochemistry tests.
Zhou
Guang-Zhou
GZ
State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Graduate School of the Chinese Academy of Sciences, Wuhan, China.
Li
Zheng-Qiu
ZQ
Zhang
Qi-Ya
QY
D000818:Animals; D000911:Antibodies, Monoclonal / Q000145:classification / Q000276:immunology*; D000914:Antibodies, Viral / Q000145:classification / Q000276:immunology*; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000302:isolation & purification / Q000378:metabolism; D045744:Cell Line, Tumor; D003593:Cytoplasm / Q000378:metabolism / Q000821:virology; D005412:Flatfishes / Q000821:virology*; D005432:Flounder / Q000378:metabolism; D005434:Flow Cytometry; D007074:Immunoglobulin G / Q000276:immunology; D007075:Immunoglobulin M / Q000276:immunology; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D012209:Rhabdoviridae / Q000276:immunology / Q000502:physiology*; D018353:Rhabdoviridae Infections / Q000821:virology; D013997:Time Factors; D014764:Viral Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D014764:Viral Proteins