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Characterization and application of monoclonal antibodies against turbot (Scophthalmus maximus) rhabdovirus
Zhou, Guang-Zhou; Li, Zheng-Qiu; Zhang, Qi-Ya; Zhang, QY, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
2006-12-01
Source PublicationVIRAL IMMUNOLOGY
ISSN0882-8245
Volume19Issue:4Pages:637-645
AbstractFive 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.; 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.
SubtypeArticle
KeywordHematopoietic Necrosis Virus Hemorrhagic Septicemia Virus Complete Genomic Sequence Scallop Chlamys-farreri Flow-cytometry Paralichthys-olivaceus Fish Rhabdovirus Spring Viremia Disease-virus Marine Fish
DepartmentChinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
Subject AreaImmunology ; Virology
DOI10.1089/vim.2006.0049
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Indexed BySCI
Language英语
WOS Research AreaImmunology ; Virology
WOS SubjectImmunology ; Virology
WOS IDWOS:000243346600006
WOS KeywordHEMATOPOIETIC NECROSIS VIRUS ; HEMORRHAGIC SEPTICEMIA VIRUS ; COMPLETE GENOMIC SEQUENCE ; SCALLOP CHLAMYS-FARRERI ; FLOW-CYTOMETRY ; PARALICHTHYS-OLIVACEUS ; FISH RHABDOVIRUS ; SPRING VIREMIA ; DISEASE-VIRUS ; MARINE FISH
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Cited Times:13[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/8730
Collection期刊论文
Corresponding AuthorZhang, QY, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
AffiliationChinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
Recommended Citation
GB/T 7714
Zhou, Guang-Zhou,Li, Zheng-Qiu,Zhang, Qi-Ya,et al. Characterization and application of monoclonal antibodies against turbot (Scophthalmus maximus) rhabdovirus[J]. VIRAL IMMUNOLOGY,2006,19(4):637-645.
APA Zhou, Guang-Zhou,Li, Zheng-Qiu,Zhang, Qi-Ya,&Zhang, QY, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China.(2006).Characterization and application of monoclonal antibodies against turbot (Scophthalmus maximus) rhabdovirus.VIRAL IMMUNOLOGY,19(4),637-645.
MLA Zhou, Guang-Zhou,et al."Characterization and application of monoclonal antibodies against turbot (Scophthalmus maximus) rhabdovirus".VIRAL IMMUNOLOGY 19.4(2006):637-645.
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