中华鲟和达氏鳇血清免疫球蛋白的木瓜蛋白酶水解片段分析

Analysis of Papain Hydrolyzed Fragments of Immunoglobulin in Serum of Chinese Sturgeon Acipenser sinesis Gray and Huso Sturgeon Georgi

首先采用饱和硫酸氨分步沉淀和Sephadex G-200凝胶层析的方法,获得了非免疫状态下中华鲟(Acipenser sinensis Gray)和达氏鳇(Huso dauricus Georgi)的血清免疫球蛋白(Ig),在此基础上使用木瓜蛋白酶水解对所获得的免疫球蛋白片段进行了酶解,并采用SDS-PAGE和Western-blot等方法分析了所获得的水解片段。结果显示,2种鲟鱼的免疫球蛋白均可被木瓜蛋白酶水解蛋白,通过Sephadex G-100凝胶层析后均可得到两个完全分离的、均一的蛋白峰。SDS-PAGE检测两个水解片段的相对分子量分别为44KD和66KD,West-ern-blot检测结果显示,66KD的片段可以在硝酸纤维素杂交膜上被各自的兔抗鲟IgM多克隆抗体所识别,而44KD片段的检测结果为阴性。这表明,2种鲟科鱼类的木瓜蛋白酶水解特性相同,提示鲟科鱼类的免疫球蛋白在免疫学及生化方面具有较大的相似性。

Saturation ammonium sulfate sedimentation and Sephadex G-200 gel chromatography were used to purify Ig from serum of Chinese sturgeon Acipenser sinesis Gray and Huso sturgeon Huso dauricus Georgi respectively,and hydrolyzed fragments by papain were analyzed.The results showed that Ig of the two fish species can be hydrolyzed into two fragments with molecule weight of 66KD and 44 KD determined by SDS-PAGE after Sephadex G-100 gel filtration.In the western blotting detection,66KD fragments can be identified by polyclonal antibody of rabbit anti sturgeon in colloxylin,while 44KD fragments cannot be deteced.A conclusion can be drawn that the hydrolyzed characters of Ig of the two fish species are same and Ig of Acipenser may have similarity in Biochemistry and Immunology.