乌鳢血清免疫球蛋白IgM的分离纯化及其兔抗血清的制备  

Purification of Serum Immunoglobulin from Argus Snakehead Fish(Ophicephalus argus) and Preparation of Rabbit Polyclonal Antibody for Argus Snakehead Fish

作　　者：林树柱[1] 李万波[1] 刘先菊[1] 杨帆[1] 全雄志[1] 全志云[1] 秦川[1] 张连峰[1] 

机构地区：[1]中国医学科学院实验动物研究所北京协和医学院比较医学中心,北京100021

出　　处：《中国比较医学杂志》2007年第11期648-651,共4页Chinese Journal of Comparative Medicine

基　　金：社会公益研究专项(2005DIA1J095)

摘　　要：目的分离纯化乌鳢血清免疫球蛋白,并制备其兔抗血清。方法用Protein A亲和层析的方法纯化乌鳢血清免疫球蛋白,通过SDS-聚丙烯酰胺凝胶电泳检测蛋白的纯度,测定其重链、轻链的分子量,免疫大耳白兔制备抗血清,利用免疫双扩散检测抗血清的效价。结果纯化了乌鳢血清免疫球蛋白,SDS-PAGE测定其重链和轻链的相对分子质量分别为78×103和27×103左右,免疫双扩散法测定兔抗乌鳢免疫球蛋白抗血清效价为1∶32。结论成功纯化了乌鳢免疫球蛋白,制备了兔抗乌鳢IgM抗血清,为研究乌鳢的免疫机制、建立乌鳢的血清学检测系统奠定了基础。Objective To purify the immunoglobulin from argus snakehead fish, and to prepare the rabbit polyclonal anti-sera. Methods Argus snakehead fish immunoglobulin was purified using staphylococcal protein A affinity chromatography. The products were examined by SDS-PAGE electrophoresis. Rabbit anti-sera were prepared by repeatedly immunizing rabbits with the purified IgM. And anti-argus snakehead fish IgM antibodies in rabbit sera were detected by agarose double immunodiffusion. Results Immunoglobulin purified from argus snakehead fish had only two bands in SDS-PAGE, and the molecular weight of heavy chain and light chain were about 78 × 10^3 and 27 × 10^3 respectively. The titer of rabbit anti-sera obtained was up to 1：32 examined by agarose double immunodiffusion. Conclusions Argus snakehead fish immunoglobulin was successfully purified, and rabbit antisera were prepared. That might provide a basis for the study on immune mechanism and developing the serological test system of Argus snakehead fish.

关 键 词：乌鳢 免疫球蛋白 亲和层析 

分 类 号：R392[医药卫生—免疫学]