蛋鸡血清载脂蛋白AⅡ的分离纯化及其抗血清的制备

Isolation of Laying Hen Serum Apolipoprotein AⅡ and Preparation of Its Specific Antiserum

本文介绍了一种在普通实验室条件下,分离纯化蛋鸡血清载脂蛋白AⅡ及其抗血清制备的一种简便易行的方法。用磷钨酸镁沉淀分离出蛋鸡血清HDL。乙醇/丙酮(1∶1V/V)脱脂后经DEAE Sepharose CL-6B和Sephadex G-150柱层析。获得的apoAⅡ经15%的尿素-SDS-PAGE电泳显示为单一条带。分子量测算大约为8673u。本方法的优点是通过使用易行的磷钨酸镁沉淀法代替了超速离心的复杂过程。不需特殊仪器,缩短了制备时间且分离效果理想。

Apolipoprotein AⅡ of laying hen serum was isolated, purified and primarily characterizated in the common laboratory. HDL from laying hen was isolated and purified from pooled serum by precipitating with phosphotungstic acid-Mg. Apolipoprotein AⅡ was obtained from HDL which was delipitated with Ethanol/ Acetone （V：V 1：1）and permeated chromatography on DEAE sepharose CL-6B column and sephadex G-150 column, primarily characterizated by 15% Urea-SDS-PAGE. The purified apolipoprotein AⅡ show only one band on Urea-SDS-PAGE and the estimated molecular weight is 8673ku.The titer of the prepared specific antiserum was 1：8, it show single sedimentary band on immune electrophoresis and double immunodifusion. The advantage of this method is that by precipating with phosphotungstic acid-Mg instead of uhracentrifugation, the special instrument, is not required, and but a satisfied result is got with short time.