兔血清ApoAI亚类的研究(Ⅱ兔血清ApoAI的纯化、鉴定及抗血清制备)

Studies of Apolipoprotein A1 Subforms in Rabbit Serum Ⅱ. The Isolation, Purification and Antiserum Preparation of Rabbit Serum Apolipoprotein A1

超速离心制备的HDL经甲醇/氯仿法脱脂后、Sephadex G-200凝胶过滤和制备电泳得到兔血清ApoAI,经SDS-PAGE、免疫电泳证明为单一蛋白质,测得MW为26786.7±297.6u,AA组份分析与文献报道一致。以此ApoA Ⅰ作为抗原,免疫豚鼠,获得抗血清。经免疫双扩散鉴定,与兔ApoAⅡ、LDL及人ApoAI无交叉反应,其效价为1:32。表明我们制备的抗血清是单价的。

Rabbit serum HDL was prepared by ultracentifugation and delipidized with methanol/chloroform. Apo-HDL was applied to Sephadex G—200 column and eleetro-phoresis to obtain apo Al. The purified apo Al was shown to be a single band by SDS—PAGE and immunoelectro-phoresis and its calculated molecular weight was 26 786.7±297.6u (mean±SD,n=6) The amino acid analysis agreed closely with that obtained by Parscan et al. The purified apo Al was used to immunize guinea pig and the sepecific antiserum to rabbit apo Al (efficiency 1:32)was collected.