摘要
IgGPH_7经木瓜蛋白酶消解产生Fab和Fc,在FPLC仪上,分别用MonoQ,Superose_(12),MonoS柱提纯Fab。提纯的Fab经还原和烷基化后,再用琥珀酸酐修饰,然后用Mono Q柱分离得到轻链和重链。最后通过SDS凝胶电泳和N端顺序测定,以鉴定它的纯度和轻、重链。
Digestion of IgGPH7 by the enzyme papain produced Fab fragments and Fc fragments.The Fab were purified by Mono Q column, Superose 12 column and Mono S column respectively on the FPLC equipment. Then the Light Chain and the Heavy Chain from the Fab were separated by Mono Q column after purified Fab were reduced, alkylated and modified. Separated Light Chain and Heavy Chain of Fab were identified by SDS-PAGE and determination of the N-terminal amino acid sequence .
