正己酸—低温乙醇沉淀法纯化小鼠腹水IgM类单抗的研究  

Purification of immunoglobulin M from mouse ascites by caproic acid and cold ethanol precipitation

作　　者：赵彭花[1] 李研[1] 王鑫[1] 赵向绒[1] 高锦伟[1] 张群英[1] 李元[1] 胡军[1] 

机构地区：[1]陕西省人民医院中心实验室,陕西西安710068

出　　处：《阜阳师范学院学报（自然科学版）》2012年第4期28-32,共5页Journal of Fuyang Normal University(Natural Science)

基　　金：2010年粤港关键领域重点突破项目东莞专项(2011A090200002)资助

摘　　要：用不同有机酸–低温乙醇纯化腹水,确定纯化效果最好的有机酸为正己酸,纯化的IgM纯度可达95%,收率将近90%;用正己酸沉淀法配合低温乙醇沉淀法分别纯化不同抗原免疫的三种IgM型单抗,对多数IgM类抗体纯度可达到85%以上,IgM回收率大于85%,表明这种纯化方法适用于多数IgM类单抗的纯化;用硫酸铵沉淀法及正己酸–低温乙醇沉淀法纯化相同的抗体,对活性都不影响,但正己酸–低温乙醇沉淀法纯化的抗体纯度远高于硫酸铵沉淀法。因此正己酸–低温乙醇沉淀法是一种简单、快速、温和的从小鼠腹水中纯化IgM方法,可满足一般实验的要求。It describes a separation of IgM from mouse ascites fluid by precipitation of caproic acid - cold ethanol precipitation method（CACE） and its effect was investigated. The results show that many other proteins are removed from serum by precipitati- on of caproic acid which is better then other. And a purity of 95% and yield almost to 90% is obtained followed by cold ethanol pre-cipitation method. Purification three IgM antibodies to different antigens with CACE the results show that the purity of the majority of IgM antibody can be up to above 85% and yield more than 80%, which means it is applicable to most of the mouse ascites fluid of IgM. Combination of the ammonium sulfate precipitation method （AS） and the CACE, the result of enzyme linked immunosorbent assay（EL1SA） indicates that any of the active of the IgM almost has no reduction compared with ascites. However, the purity of the IgM purified by CACE is much higher than the lgM purified by AS. This represents a simple, rapid and gentle approach to isolate lgM from mouse ascites fluid and can meet the general experimental requirements.

关 键 词：免疫球蛋白M(IgM) 纯化 正己酸 低温乙醇 小鼠腹水 

分 类 号：R392.11[医药卫生—免疫学]