人血清氧化低密度脂蛋白自身抗体的提取和初步鉴定

Isolation and identification of human anti-oxidized LDL autoantibodies

目的提取和鉴定人血清氧化低密度脂蛋白(Ox-LDL)自身抗体。方法采用溴化氰活化的Sepharose4B交联Ox-LDL制备免疫吸附层析柱,从正常人血清中提取抗Ox-LDL自身抗体,采用ELISA法鉴定其免疫反应性,散射免疫比浊法分析自身抗体种类。结果用NaHCO3、醋酸盐和KCNS溶液洗脱,分别收集洗脱峰。8份正常人血清中均收集到Ox-LDL自身抗体,与Ox-LDL均具有良好的反应性。自身抗体种类中以IgG为主,其次为IgM。结论此法提取的抗Ox-LDL自身抗体同Ox-LDL具有良好的反应能力。

Objective To isolate and identify human anti-oxidized low density lipoprotein （LDL） autoantibodies. Methods Oxidized LDL （Ox-LDL） autoantibodies were isolated from eight healthy subjects by affinity chromatography with using an Ox-LDL cross-linked Sepharose 4B column. Results Three elution peaks of Ox-LDL autoantibodies were washed out from the Ox-LDL column by NaHCO3, acetate and KCNS buffer in turn and collected respectively. Ox-LDL autoantibodies were isolated from all the selected healthy subjects. All the three elution peaks had good reactivity with Ox-LDL. Among them,the elution peaks by KCNS buffer had high protein level and strong reactivity with Ox-LDL. The types of these antibodies included IgG and IgM, and IgG was the main type. Conclusion The isolated Ox- LDL autoantibodies had strong reactivity with Ox-LDL.