Miekka S I, Ingham K C, Menache D
Thromb Res. 1982 Jul 1;27(1):1-14. doi: 10.1016/0049-3848(82)90272-9.
Simplified procedures have been developed for isolation of human plasma fibronectin by affinity chromatography on gelatin-agarose. In one method, fibronectin is eluted with 3 M urea, and this reagent is quickly removed by adsorbing the protein onto heparin-agarose, followed by 0.4 M NaCl elution. In a shorter process, fibronectin is eluted from gelatin-agarose simply by decreasing the buffer pH below 6. After lyophilization the purified protein can be readily dissolved in water. The fraction not adsorbed to gelatin can be used to purify other proteins, including factor VIII whose procoagulant activity is quantitatively recovered.
已开发出通过在明胶 - 琼脂糖上进行亲和色谱法分离人血浆纤连蛋白的简化程序。在一种方法中,用3M尿素洗脱纤连蛋白,然后通过将蛋白质吸附到肝素 - 琼脂糖上快速去除该试剂,接着用0.4M NaCl洗脱。在一个更简短的过程中,只需将缓冲液pH降至6以下即可从明胶 - 琼脂糖上洗脱纤连蛋白。冻干后,纯化的蛋白质可很容易地溶解于水中。未吸附到明胶上的部分可用于纯化其他蛋白质,包括其促凝血活性可定量回收的凝血因子VIII。