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.
出处
《中国生化药物杂志》
CAS
CSCD
北大核心
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)
关键词
红细胞凝集素
分离纯化
甲状腺球蛋白
共沉淀
erythrohemagglutinin（PHA-E）
purification
thyroglobulin
co-precipitation
