应用血清分离试剂盒纯化血清标志蛋白的研究

Study on purification of protein from human serum with kit-serum fractionation

目的:研究采用血清分离试剂盒初步分离血清标志蛋白的效果。方法:应用血清分离试剂盒对血清中标志蛋白进行梯度pH洗脱,根据目的蛋白的洗脱条件,选择适合的缓冲液进行离子交换层析分离。结果:WCX-2和IMAC-3-Cu芯片检测结果显示,血清经血清分离试剂盒梯度pH洗脱后可得到初步分离,各组分中的蛋白数量都比原血清明显减少。其中在第3、4组分分别检测到目的蛋白M7789.8和M4300.80。选择缓冲液(20 mmolNaAc,pH4.0)进行离子交换层析,成功分离到目的蛋白M7789.8和M4300.80。结论:应用血清分离试剂盒不仅可以对血清进行初步分离,还能了解到目的蛋白表面一些氨基酸的基本构成和等电点范围,为后续进一步分离纯化目的蛋白提供参考。

Objective：To study the effect of separating protein from human serum with kit-serum fractionation.Methods：Protein in human serum was eluted by the kit-serum fractionation according to different pH values.The suitable buffer solution selected to use on the ion-exchange chromatography was based on the eluting condition of aim protein.Results：The results of WCX-2 and IMAC-3-Cu protein chips showed that serum was separated preliminarily after serum was eluted by the kit-serum fractionation according to different pH values.The number of protein in each fraction was less than the original serum.The aim protein M7 789.8 in the third fraction and M4 300.80 in the fuorth fraction were detected respectively and separated by the ion-exchange chromatography with buffer solution（20 mmol NaAc,pH4.0）.Conclusion：The kit-serum fractionation can not only be used to separate protein from serum preliminarily but also study the aim protein′s basic composition of some amino acid on the surface and scope of pI.It provides an important reference to separate and purify the aim protein from serum further.