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Characterization and Application of Monoclonal Antibodies against Turbot (Scophthalmus maximus) Rhabdovirus
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Characterization and Application of Monoclonal Antibodies against Turbot (Scophthalmus maximus) Rhabdovirus
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Journal Article
Characterization and Application of Monoclonal Antibodies against Turbot (Scophthalmus maximus) Rhabdovirus
2006
Overview
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.
Publisher
Mary Ann Liebert, Inc,SAGE Publications,Mary Ann Liebert
Subject
/ Antibodies, Monoclonal - classification
/ Antibodies, Monoclonal - immunology
/ Antibodies, Viral - classification
/ Antibodies, Viral - immunology
/ Antigens, Viral - isolation & purification
/ Antigens, Viral - metabolism
/ Immunoglobulin G - immunology
/ Immunoglobulin M - immunology
/ Mice
