美洲黑石斑鱼血清IgM纯化及其兔抗血清部分特性

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

摘要:

采用蛋白 A 亲和层析法对健康美洲黑石斑鱼血清中的免疫球蛋白(IgM)进行分离纯化。实验结果表明,蛋白 A亲和层析法一步纯化即可得到电泳纯IgM,其重链和轻链的分子量约 74.1 ku和26.0 ku。用纯化的黑石斑鱼IgM免疫实验兔，获得效价高达1∶128 000的兔抗黑石斑鱼IgM血清。Western-blot和间接ELISA 检测证明，本工作制备的兔抗美洲黑石斑鱼IgM血清具有较高的特异性，为后续的黑石斑鱼免疫学研究奠定了基础。

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.