PRODUCTION AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES AGAINST THE HAEMOCYANIN OF THE CHINESE SHRIMP, FENNEROPENAEUS CHINENSIS (OSBECK, 1765)

Two protein bands with molecular masses of 73 kDa and 75 kDa were purified from haemolymph of the Chinese shrimp, Fenneropenaeus chinensis (Osbeck, 1765), by affinity chromatography coupled with purified white spot syndrome virus, which were identified as haemocyanin subunits by mass spectrometry. Fourteen hybridomas secreting monoclonal antibodies against Chinese shrimp haemocyanin were obtained by immunization of Balb/C mice with purified haemocyanin preparation, which were selected on the basis of the indirect enzyme-linked immunosorbent assay (ELISA). Three of these, designated as 2H3, 4E5, and 1G8 were cloned by limiting dilution and characterized with western blotting. Under reducing conditions in Western blotting, all three MAbs could specifically react with 73 and 75 kDa subunits of haemocyanin of E chinensis. MAb 2H3 could also react with the native haemocyanin in haemolymph by immunoprecipitation, and cross-react with the haemocyanins of Marsupenaeus japonicus (Bate, 1888), Litopenaeus vannamei (Boone, 1931) and Procambarus clarkii (Girard, 1852).