Purification of IgM from black sea bass (Centropristis striata) and characterization of rabbit antiIgM serum

Abstract:

The purification of serum immunoglobulin in Centropristis striata was carried out by using the protein Asepharose affinity chromatography and the final purified product was examined by SDSPAGE electrophoresis. The experimental results showed that IgM with a high purity could be prepared after one purifying step of the protein Asepharose affinity chromatography. SDSPAGE showed that its heavy chain and light chain were 74.1 ku and 26 ku respectively. Serum against IgM from Centropristis striata was prepared by repeatedly immunizing New Zealand rabbits with the purified IgM and the titers of antibodies reached 1∶128 000. A high specificity of the obtained antiserum was qualified by indirect ELISA and Western Blot assays, which could supply a necessary approach in the following immunological studies for Centropristis striata.