Rastogi S C, Clausen J
Clin Chim Acta. 1980 Feb 14;101(1):85-92. doi: 10.1016/0009-8981(80)90059-5.
A method for isolation of two multiple sclerosis(MS)-specific brain antigens [3] is described. The brain cytosol was passed through a Con A-Sepharose column and the glycoproteins containing MS-specific antigens (MSG) were eluted with 50 mmol/l methyl-alpha-D-glucopyranoside, pH 6. By fractionation of MSG on DEAE-cellulose, a fraction containing MS-specific antigens (MSG2) was eluted in 0.25 mol/l NaCl, after prior elution of other proteins with distilled water. The MS specific antigens in the MSG2 fraction were traced and characterized by crossed immunoelectrophoresis and isoelectric focusing. A quantitative determination of proteins eluted in MSG2 revealed that the 2 MS-specific antigens had been purified 10 000 times. Using this method to isolate the two MS-specific antigens of autopsy brains, they were found to be present in 7/7 MS brains and absent in 9/9 non-MS brains studied. The method described in the present communication may be used both for the partial purification of the MS-specific antigens as well as for the tracing of the specific antigens in different topographical areas of the MS brains.
本文描述了一种分离两种多发性硬化症(MS)特异性脑抗原的方法[3]。脑细胞质通过伴刀豆球蛋白A-琼脂糖柱,含有MS特异性抗原(MSG)的糖蛋白用50 mmol/l甲基-α-D-吡喃葡萄糖苷(pH 6)洗脱。通过在DEAE-纤维素上对MSG进行分级分离,在用蒸馏水预先洗脱其他蛋白质后,在0.25 mol/l NaCl中洗脱得到含有MS特异性抗原的级分(MSG2)。通过交叉免疫电泳和等电聚焦对MSG2级分中的MS特异性抗原进行追踪和鉴定。对在MSG2中洗脱的蛋白质进行定量测定表明,这两种MS特异性抗原已被纯化了10000倍。使用该方法分离尸检脑的两种MS特异性抗原,发现它们存在于所研究的7/7例MS脑中,而在9/9例非MS脑中不存在。本通讯中描述的方法可用于MS特异性抗原的部分纯化以及在MS脑不同区域追踪特异性抗原。