几种部分纯化单克隆抗体方法的比较

COMPARATIVE STUDIES OF DIFFERENT PURIFICATION METHODS FOR MONOCLONAL ANTIBODY ( McAb )

比较了几种提纯方法对流行性出血热病毒的McAb E_5腹水的提纯效果,证实辛酸—硫酸铵法最好,抗体活性的回收率可高达100％。经聚丙烯酰胺电泳分析后,只出现免疫球蛋白重链和轻链两条带,纯度较好。辛酸用量很省,每毫升腹水只和33μ1。作用条件的要求也不高,室温作用≥30分钟,4℃作用≥60分钟,离心速度≥13 000r/min,方法简便,适于推广应用.

Several purification methods were compared. The results shoved that caprylic acid clarification followed by ammonium sulfate precipitation is a simple, rapid and inexpensive method. The McAb from mouse ascitic fluid prepared by caprylic acid/ammonium sulfate was high purified. Only IgG H chain and IgG L chain can be seen by SDS-PAGE analysis of purified sample. The IF titer of purified McAb was 10,240 or as high as the original material. 33μl of caprylic acid is needed for one ml of mouse ascitic fluid McAb. This method is better than protein A-Sepharose chromatography, DEAE-32 cellulose chromatography, ammonium sulfate precipitation only, caprylic acid clarification only and HPT chromatography.