美国红鱼免疫球蛋白的分离纯化  

The Purification of Serum Immunoglobulin of Sciaenops ocellatus

作　　者：罗轻蕾[1,2,3] 闫茂仓[2,3] 陈少波[2,3] 谢起浪[2,3] 单乐州[2,3] 朱进[4] 艾为明[1] 

机构地区：[1]温州医学院生命科学学院,浙江温州325005 [2]浙江省海洋水产养殖研究所,浙江温州325005 [3]浙江省近岸水域生物资源开发与保护重点实验室,浙江温州325005 [4]温州医学院附属第一医院肿瘤外科,浙江温州325005

出　　处：《科技通报》2010年第4期623-627,共5页Bulletin of Science and Technology

基　　金：浙江省科技厅科研院所专项公益技术攻关项目(2007F10009);浙江省科技厅科研院所青年人才计划项目(2006R20001);温州市科技海兴海项目(S20070051);浙江省科技厅创新团队建设与人才培养(2009F2009)

摘　　要：对灿烂弧菌灭活疫苗接种后的美国红鱼提取血清,采用硫酸铵沉淀和SephadexG-200凝胶层析结合的方法分离纯化出美国红鱼血清免疫球蛋白,并且通过SDS-PAGE对纯化蛋白的部分特性进行分析。结果表明,通过硫酸铵沉淀法可以沉淀血清中大部分的蛋白,电泳条带比较丰富,但仍然含有较多的杂蛋白,经过SephadexG-200凝胶柱层析方法分离,获得纯度较高的免疫球蛋白,经β—巯基乙醇处理后解离形成重链和轻链两个亚单位,分子量分别为69.2 kDa,29.5 kDa,表明美国红鱼血清IgM的分子量与硬骨鱼IgM的分子量范围（H链60~81kDa,L链20-30kDa）是一致。The serum was extracted from Sciaenops ocellatus which was inoculated with Vibrio splendidus .The serum immunoglobulin of Sciaenops ocellatus was separated and purificated by ammonium sulfate precipitation and SephadexG- 200 gel chromatography .And partial characteristics of purified proteins were analyzed by SDS-PAGE. The results showed that ：The most proteins in serum were precipitated by the method of ammonium sulfate precipitation,and the electrophoresis strips were very rich. But there were many hybridproteins in the serum.When the serum immunoglobulins of Sciaenops ocellatus which were purificated by ammonium sulfate precipitation and SephadexG-200 gel chromatogra- phy,it obtained the high purity of immunoglobulin.After decomposited by a β-mercaptoethanol , there were two chains in the IgM of the Sciaenops ocellatus ,that is the H chain （69.2 kD ） and L chain （29.5 kDa） . It also revealed that the Immunoglobulin M of the serum in the Sciaenops ocellatus was in line with that of the serum in the teleost which the size of H chain was from 60kDa to 81 kDa and the size of L chain was from 20 kDa to 30 kDa.

关 键 词：美国红鱼 免疫球蛋白 硫酸铵沉淀 葡聚糖SephadexG-200凝胶层析 

分 类 号：TQ937[化学工程]