【摘要】 本研究将离子交换层析和金属螯合亲和层析方法相结合,建立了系统的分离纯化β-伴大豆球蛋白天然状态亚基的方法,通过梯度脲透析复性,使纯化亚基恢复天然构象。利用β-伴大豆球蛋白免疫动物制备抗血清对纯化亚基的免疫活性进行检测。结果表明:离子交换层析结合金属螯合亲和层析方法可以有效地纯化β-伴大豆球蛋白的α、α’和β亚基,该方法产率高,制备的亚基纯度好,而且能与抗β伴大豆球蛋白血清特异性结合,表明提纯的亚基具有免疫活性。更多还原

【Abstract】 A systemic approach for purification of natural α,α’ and β subunits from soybean protein β-conglycinin was established using a combination of ion exchange chromatography and metal chelating chromatography.Then,purified subunits were returned to natural conformations via gradual dialysis against buffers containing decreased concentration of urea.Furthermore,immunological activities of purified subunits were determined with the antiserum which was prepared through immunizing rabbit with β-conglycinin.Results showed that α,α’ and β subunits can be purified effectively from β-conglycinin by ion exchange chromatography and metal chelating chromatography.The production yield by this method is high and prepared subunits are highly pure.The subunits can specifically bind to β-conglycin antiserum and have immunological activity.更多还原