Generation and characterization of monoclonal antibodies against the flounder Paralichthys olivaceus rhabdovirus

• Published in: Journal of Virological Methods
• Main Authors: Zhou, Guang-Zhou, Gui, Lang, Li, Zheng-Qiu, Yuan, Xiu-Ping, Zhang, Qi-Ya, Zhang, QY, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
• Format: Article in Journal/Newspaper
• Language: English
• Published: 2008
• Subjects: Fish Rhabdoviruses; Monoclonal Antibodies; Immunocytochemistry; Flow Cytometry; Biochemical Research Methods; Biotechnology & Applied Microbiology; Virology; Science & Technology; Life Sciences & Biomedicine; Biochemistry & Molecular Biology; VIRAL HEMORRHAGIC SEPTICEMIA; SCOPHTHALMUS-MAXIMUS RHABDOVIRUS; LYMPHOCYSTIS DISEASE VIRUS; COMPLETE GENOMIC SEQUENCE; FISH RHABDOVIRUS; SPRING VIREMIA; CARP VIRUS; RABIES; ASSAY; DIAGNOSIS; Scophthalmus maximus
• Online Access: http://ir.ihb.ac.cn/handle/152342/8178; https://doi.org/10.1016/j.jviromet.2007.11.010

Two MAbs (3C7 and 3C9) against flounder Paralichthys olivaceus rhabdovirus (PORV) were generated with hybridoma cell fusion technology and characterized by an indirect enzyme-linked immunosorbent assay, isotype test, Western blot and immunodot analysis and immunofluorescence assay. Isotyping tests demonstrated that both of the two MAbs belonged to IgM subclass. Western blot analysis showed the MAbs reacted with 42, 30, and 22 kDa viral proteins, which were localized within the cytoplasm of PORV-infected grass carp ovary (GCO) cells analyzed by indirect immunofluorescences tests. The MAb 3C7 was also selected at random for detecting virus antigens in the inoculated grass carp tissues by immunohistochemistry assay. Flow cytometry tests showed that at the 36 h postinfection (0.25 PFU/cell), the 23% PORV-infected GCO cells could be distinguished from the uninfected cells with the MAb 3C7. Such MAbs could be useful for diagnosis and potential treatment of viral infection. (C) 2007 Elsevier B.V. All rights reserved. Two MAbs (3C7 and 3C9) against flounder Paralichthys olivaceus rhabdovirus (PORV) were generated with hybridoma cell fusion technology and characterized by an indirect enzyme-linked immunosorbent assay, isotype test, Western blot and immunodot analysis and immunofluorescence assay. Isotyping tests demonstrated that both of the two MAbs belonged to IgM subclass. Western blot analysis showed the MAbs reacted with 42, 30, and 22 kDa viral proteins, which were localized within the cytoplasm of PORV-infected grass carp ovary (GCO) cells analyzed by indirect immunofluorescences tests. The MAb 3C7 was also selected at random for detecting virus antigens in the inoculated grass carp tissues by immunohistochemistry assay. Flow cytometry tests showed that at the 36 h postinfection (0.25 PFU/cell), the 23% PORV-infected GCO cells could be distinguished from the uninfected cells with the MAb 3C7. Such MAbs could be useful for diagnosis and potential treatment of viral infection. (C) 2007 Elsevier B.V. All rights reserved.