亲和共沉淀从红芸豆中提取红细胞凝集素及其性质研究  

Purification and partial characterization of erythrohemagglutinin from Phaseolus vulgaris by co-precipitation

作　　者：张涛[1] 杨雨晗[2] 何涛[3] 廖涛[3] 张芹[3] 吴昌英[3] 

机构地区：[1]成都医学院生物医学系生物技术实验室,四川成都610083 [2]成都医学院生物医学系细胞生物学和遗传学教研室,四川成都610083 [3]成都医学院生物医学系生物技术专业,四川成都610083

出　　处：《中国生化药物杂志》2011年第3期202-205,共4页Chinese Journal of Biochemical Pharmaceutics

基　　金：四川省教育厅科研基金项目(10ZC090)

摘　　要：目的研究一种从红芸豆中提取红细胞凝集素(PHA-E)的新方法。方法红芸豆经过粗提,然后和甲状腺球蛋白的粗提取液进行共沉淀,利用超滤制得PHA-E产品,采用电泳和质谱进行鉴定,并研究其凝血活力。结果所获样品的纯度高,其亚基相对分子质量为32 000,等电点为6.5。经质谱鉴定,纯化产品为PHA-E。使兔红细胞50%凝集的PHA-E的最低浓度为2.45μg/mL,单糖不影响PHA-E凝血活力,EDTA抑制其凝血活力,Zn2+促进其凝血。结论通过亲和共沉淀的方法能快速简便的提取活力高的PHA-E。Purpose To study new techniques of the separation and purification of erythrohemagglutinin（PHA-E） from Phaseolus vulgaris.Methods PHA-E was purified from Phaseolus vulgari extract by co-precipitation with thyroglobulin extract and by ultrafiltration.The purity was identified by electrophoresis and spectrometry.The ability of agglutination was determined with 2% erythrocytes of rabbit.Results SDS-PAGE revealed that PHA-E has apparent subunit molecular weights of 32 kD and the PAGE revealed that PHA-E has a single outline.The isoelectric point of the PHA-E was 6.5.Identifications by spectrometry demonstrated that the product was PHA-E.It can promote agglutination of rabbit erythrocytes at 2.45 μg/mL but its activity was not inhibited by any monosaccharide.EDTA can inhibit the ability of agglutination,and Zn^2＋ can accelerate it.Conclusion Purified PHA-E can be obtained by co-precipitation with thyroglobulin.

关 键 词：红细胞凝集素 分离纯化 甲状腺球蛋白 共沉淀 

分 类 号：R284.2[医药卫生—中药学] R285.5[医药卫生—中医学]