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纤维连结蛋白(Fibronectin,FN)是一种分子量为44万的糖蛋白。本文报道应用溴化氰活化琼脂糖珠4B与明胶偶联亲和层析柱来制备FN的方法。以健康男性血浆为原料,加入层析柱中,使FN吸附在明胶上,用0.1mol/L三羟甲基氨基甲烷(Tris)—盐酸(HCI)缓冲液(pH7.5,含0.02mol/L柠檬酸钠)冲洗,除去不结合物质。然后用含1mol/L NaCl的上述缓冲液洗杂蛋白,再用含3mol/L脲的上述缓冲液洗脱,得到高纯度的FN。此方法简单,所得FN纯度高,效果好。
Fibronectin (FN) is a glycoprotein with a molecular weight of 440,000. This article reports the preparation of FN using cyanogen bromide activated Sepharose 4B with gelatin-coupled affinity chromatography column. Using healthy male plasma as raw material, the FN was added to the column to adsorb FN on the gelatin. The FN was adsorbed on the gelatin with 0.1 mol / L Tris-HCl (pH 7.5) containing 0.02 mol / L sodium citrate) to remove non-binding substances. Then, the protein was washed with the above buffer containing 1 mol / L NaCl and then eluted with the above buffer containing 3 mol / L of urea to obtain high purity FN. This method is simple, the resulting FN high purity, good effect.