【摘要】 本研究建立了一种在普通实验室条件下,分离纯化鸡血清载脂蛋白AⅡ(apo AⅡ)及其抗血清制备的方法。用磷钨酸镁沉淀法分离出鸡血清高密度脂蛋白(HDL),乙醇/丙酮(1∶1)脱脂后经DEAE Sepharose CL-6B和Sephadex G-150柱层析,获得的apo AⅡ经尿素-SDS-PAGE电泳显示为单一条带,相对分子质量为8 673。采用弗氏佐剂充分乳化的上述蛋白对新西兰雄兔进行多次多点皮内或皮下注射,分离兔抗鸡apo AⅡ血清后自行制备标准血清(1.112 g/L),采用免疫透射比浊法测得鸡血清apo AⅡ的生理含量为(1.295±0.21)g/L。此方法简便易行、准确可靠,有利于临床推广应用。更多还原

【Abstract】 A convenient method of purifying and identifying apolipoprotein AⅡ from chicken serum in common laboratory was established in this study.HDL in chicken was isolated and purified from pooled serum by precipitating with phosphotungstic acid-Mg,then apolipoprotein AⅡ was obtained from HDL which was delipitated with ethanol/acetone(1∶1)and permeated chromatography on DEAE sepharose CL-6B column and sephadex G-150 column.The purified apolipoprotein AⅡ showed only one band on Urea-SDS-PAGE and the estimated molecular weight was 8 673.To obtained the specific antiserum,New Zealand male rabbit was injected subcutaneously with the apolipoprotein AⅡ protein which was fully emulsified with equal volume of Freund’s adjuvant for many times,after the antiserum were obtained from the rabbits blood,the standard serum was prepared(1.112 g/L).Subsequently,the physiological concentration of apo AⅡ in this breed chicken which was(1.295±0.21) g/L were measured by immunoturbidimetric assay.This method was proved to be accurate,reliable and can be used in clinical practice.更多还原