利用国产IVIg分离纯化抗-Aβ初探  

The isolation and purification of anti-Aβ antibody from domestic IVIg through affinity chromatography

作　　者：辛叶 曹海军[1] 叶生亮[1] 李长清[1] 

机构地区：[1]中国医学科学院北京协和医学院输血研究所,四川成都610052

出　　处：《中国输血杂志》2015年第5期520-523,共4页Chinese Journal of Blood Transfusion

摘　　要：目的利用国产IVIg初步探讨抗-Aβ的分离纯化技术。方法首先,用人工合成的人源Aβ1-42进行聚合以制备Aβ寡聚体并通过SDS-PAGE对其进行鉴定。其次,通过偶联缓冲液(0.6 mol/L柠檬酸钠、0.2 mol/L碳酸钠,PH:10)将制备好的Aβ偶联到Ultra Link Biosupport凝胶上,然后将稀释后的IVIg加入到偶联好的凝胶上,经过平衡(0.1 mol/L PBS,PH:7.4)、洗涤(0.1 mol/L PBS,PH:7.4)、洗脱(0.1 mol/L甘氨酸缓冲液,PH:2.2),从而分离纯化出抗-Aβ。最后,通过ELISA和SDS-PAGE鉴定纯化出的抗-Aβ抗体。结果聚合的Aβ1-42除了含有少量尚未聚合的单体外,主要为不同分子量的寡聚体(如:三聚体、八聚体、十二聚体等)。所获得抗-Aβ回收率为21.1%、比活为9 671.9 ng/mg、提纯倍数为248。结论使用离心柱法亲和层析可从国产IVIg制品中分离纯化出抗-Aβ。Objective To study the isolation and purification of anti-Aβ antibody from domestic IVIg. Methods Aβoligomers were first prepared from human synthetic Aβ1- 42 and then the oligomers were analyzed by SDS-PAGE. Second,Aβ was coupled to UltraL ink Biosupport resin by coupling buffer（ 0. 6 mol / L sodium citrate,0. 2 mol / L sodium carbonate,PH： 10）,and then diluted IVIg was loaded to the coupled resin. Anti-Aβ antibody was purified through the following proce-dure： balance（ 0. 1mol / L PBS,PH： 7. 4）,wash（ 0. 1 mol / L PBS,PH： 7. 4） and elute（ 0. 1 mol / L glycine buffer,PH： 2.2）. Finally,Aβ specific antibody was identified by ELISA andSDS-PAGE. Results In addition to discovery of several monomers,low molecular weight and high molecular weight in Aβoligomers indicated the presence of trimer,octamer and dodecamer. The rate,specific activity and purification level of Aβspecific antibody in eluate were 21. 1%,9671. 9 and 248,respectively. Conclusion Anti-Aβ antibody could be purified from domestic IVIg through affinity chromatography with centrifuge column.

关 键 词：阿尔茨海默病 静注人免疫球蛋白 Β淀粉样蛋白 离心柱 亲和层析 

分 类 号：R818.052[医药卫生—放射医学] R592[医药卫生—临床医学]