黄鳝血清白蛋白的纯化及抗体制备

Purification and antibody preparation of serum albumin from Monopterus albus

通过阴离子交换层析和凝胶过滤层析,从黄鳝血清中得到其血清白蛋白,命名为MAA-serum,分子量为67kDa,为单链非糖蛋白,其N末端氨基酸序列为GHKKW。用0.5mg/只MAA-serum作为免疫源免疫大白兔,经3次免疫后,抽取大白兔血液,分离血清,饱和硫酸氨沉淀,获得抗MAA-serum的多克隆抗体。经ELISA方法测定该抗体与黄鳝血清白蛋白的免疫反应,抗体滴度为1∶100000。经初步研究发现黄鳝皮肤中也有大量血清白蛋白分布,该抗体为进一步研究MAA-serum在皮肤中的分布及生物学功能打下基础。

Serum albumin of Monopterus albus was obtained through the steps of mixing ion-exchange and gel filtration chromatography, and named MAA-serum, which is a single chain protein with molecular weigh of 67 kDa. MAA-serum is not a glycoprotein. The N-terminal amino acid sequence is GHKKW. The purified MAA-serum was used to immunize rabbits to prepare polyclonal antibody with the dosage of 0.5 mg/per rabbit. After immunized three times, the rabbits were anesthetized with ether, then the blood was collected by a cardiac puncture, and precipitate with saturated ammonium sulfate to obtain polyclonal antibody. ELISA data demonstrated that the antibody titer of the serum was up to 1：100 000. The anti MAA-serum polyclonal antibody had a high quality of potency and specificity. The antibody provided an useful experimental tool to profoundly research the tissue expression profile and biological function of MAA-serum.