大豆凝集素的分离纯化及活性鉴定  

Isolation and Purification and Active Identification of Soybean Agglutinin

作　　者：车东升[1] 刘飞飞[1] 穆成龙[1] 张海全[1] 孙泽威[1] 秦贵信[1] 

机构地区：[1]吉林农业大学动物生产及产品质量安全教育部重点实验室,动物科学技术学院,长春130118

出　　处：《吉林大学学报（理学版）》2012年第5期1041-1044,共4页Journal of Jilin University:Science Edition

基　　金：国家自然科学基金(批准号:30871802);吉林农业大学青年基金(批准号:201026)

摘　　要：以D-半乳糖胺为配基、FF-sepharose 4B为固定相,制备大豆凝集素亲和层析填料.分离和纯化大豆中的大豆凝集素蛋白,测定其纯度、免疫原性和凝集活性,并与美国Sigma公司的标准品比较.结果表明:每毫升GalN-FF-Sepharose-4B层析填料能结合10 mg大豆凝集素;纯化获得的大豆凝集素分子量为30 000;SDS-PAGE测定纯度大于98%;Western blot结果为阳性;1 mg/mL SBA血集效价为1/210;其纯度、免疫原性和凝集活性等指标与大豆凝集素标准品相同,与理论值相符.The experiment was designed to use D-galactosamine as ligand, and FF-Sepharose 4B as immobilization phase to synthesize D-galactosamine-FF-Sepharose chromatography stuffing. Soybean agglutinin was isolated and purified with its purity measured; immunogenieity and agglutination activity were determined and then compared with those of American Sigma company standard sample. The result shows the GalN-FF- Sepharose-4B could adsorb 10 mg of SBA per milliliter. After purification, soybean agglutinin molecular weight was 30 000; SDS-PAGE determined purity of it was over 98% ; the western blot test was positive; 1 mg/mL SBA hemagglutination titer was 1/210; its purity, immunogenicity and agglutination activity were thesame as those of soybean agglutinin standard sample and consistent with the theoretical values.

关 键 词：大豆凝集素 纯化 亲和层析 

分 类 号：Q5-33[生物学—生物化学]