Generation and characterization of monoclonal antibodies against the flounder Paralichthys olivaceus rhabdovirus.
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.
Zhou
Guang-Zhou
GZ
State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.
Gui
Lang
L
Li
Zheng-Qiu
ZQ
Yuan
Xiu-Ping
XP
Zhang
Qi-Ya
QY
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000276:immunology* / Q000302:isolation & purification; D000956:Antigens, Viral / Q000032:analysis; D015153:Blotting, Western; D002347:Carps; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D005393:Fish Diseases / Q000821:virology*; D005412:Flatfishes / Q000821:virology*; D007075:Immunoglobulin M / Q000276:immunology / Q000302:isolation & purification; D007668:Kidney / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D012209:Rhabdoviridae / Q000276:immunology*; D018353:Rhabdoviridae Infections / Q000821:virology*; D013154:Spleen / Q000821:virology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D007075:Immunoglobulin M