This work was to prepare and characterize a monoclonal antibody against MCP COE protein of Chinese giant salamander (Andrias davidianus) iridovirus (CGSIV), so as to provide a valuable tool for CGSIV diagnosis and further researches on CGSIV pathogenesis. After being purified by nitrilotriacetic acid (Ni-NTA) agarose resin, the recombinant MCP COE of CGSIV was used to immunize the BALB/c mice. Three days after the last immunization, spleen cells were removed and fused with myeloma cells. One hybridoma cell strain against the recombinant protein of MCP COE was obtained by screening with the indirect ELISA and limiting dilution assays, which was named 1M6. The McAb of 1M6 was identified to be IgG2b isotype with κ light chain and showed specific reactivity with CGSIV and the recombinant protein of MCP COE. The McAb was largely produced in mouse ascites and then depurated with saturated ammonium sulfate. Indirect ELISA assay showed that the valences of the McAbs in the cells culture supertanant and in mouse ascites were 1:1600 and 1:2×10⁶ respectively. Affinity constant of the McAb of 1M6 was 2×10⁵. By immunofluorescence assay, the presence of CGSIV was observed in not only the cytoplasm but also the nucleus of CGSIV infected EPC cells. In conclusion, the prepared McAb has high titer and good specificity, which will lay the foundation for further studies on the control of CGSIV.