用膜渗滤亲和层析法纯化腹水单克隆抗体(McAb).采用硝酸纤维素膜(NCM)作固相支持物吸附抗原,用负压使小鼠腹水渗滤NCM,在滤过的同时腹水中的McAb不断结合于NCM上吸附的抗原,再将McAb从NCM上解离,从而得到高纯度的McAb.用此法纯化抗白蛋白(Alb)McAb.结果提纯的Alb McAb纯度达PAGE电泳单条带,将此McAb点样NCM用于斑点免疫渗滤法(DIFA)检测Alb,其灵敏度比用腹水点样时高20倍.该法快速简便,可代替亲和层析柱用于纯化McAb.
A new way for affinity chromatography has been developed to purify the McAb in ascites. Using nitrocellulose membrane(NCM) as solid-phase to adsorb the antigen, filtrating the sample under negative pressure through the NCM which could bind the relevant antibody to the antigen adsorbed on, then dissociating the antibody from the NCM in the purified form, this is the process of the so called membrane filtration affinity chromatography(MFAC). The albumin(Alb) McAb in ascites has been purified with the MFAC. The purified McAb showed a single band in the polyacrylamide gel electrophoresis(PAGE).While used in dot immunofiltration assay(DIFA) to detect Alb standard, the sensibility of the purified McAb was 20 times more than that of the ascites. The simple and effective MFAC can be used as a new affinity chromatography to purify antibodies in ascites.
徐兵,朱忠勇,唐玉钗,兰小鹏.膜渗滤亲和层析法纯化腹水单克隆抗体[J].生物化学与生物物理进展,1997,24(4):373-375
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