大豆凝集素的分离纯化及活性鉴定

J4 ›› 2012, Vol. 50 ›› Issue (05): 1041-1044.

大豆凝集素的分离纯化及活性鉴定

车东升, 刘飞飞, 穆成龙, 张海全, 孙泽威, 秦贵信

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

收稿日期:2012-01-17 出版日期:2012-09-26 发布日期:2012-09-29
通讯作者: 秦贵信 E-mail:qgx@jlau.edu.cn

Isolation and Purification and Active Identificationof Soybean Agglutinin

CHE Dongsheng, LIU Feifei, MU Chenglong, ZHANG Haiquan, SUN Zewei, QIN Guixin

1. Animal Production &|Product Quality and Security Key Lab, Ministry of Education,College of Animal Science and Technology, Jilin Agricultural
University, Changchun 130118, China

Received:2012-01-17 Online:2012-09-26 Published:2012-09-29
Contact: QIN Guixin E-mail:qgx@jlau.edu.cn

摘要/Abstract

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1/2^10; 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.

关键词: 大豆凝集素；纯化；亲和层析

Abstract:

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; immunogenicity 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 the same as those of soybean agglutinin standard sample and consistent with the theoretical values.

Key words: soybean agglutinin, purification, affinity chromatography

中图分类号:

Q5\|33

车东升, 刘飞飞, 穆成龙, 张海全, 孙泽威, 秦贵信. 大豆凝集素的分离纯化及活性鉴定[J]. J4, 2012, 50(05): 1041-1044.

CHE Dong-Sheng, LIU Fei-Fei, MU Cheng-Long, ZHANG Hai-Quan, SUN Ze-Wei, QIN Gui-Shen. Isolation and Purification and Active Identificationof Soybean Agglutinin[J]. J4, 2012, 50(05): 1041-1044.