van den Eijnden-van Raaij A J, Koornneef I, van Zoelen E J
Hubrecht Laboratory, Netherlands Institute for Developmental Biology, Utrecht.
Biochem Biophys Res Commun. 1988 Nov 30;157(1):16-23. doi: 10.1016/s0006-291x(88)80004-4.
A new method was developed for the purification of type beta transforming growth factor from human platelets. This method is a three-step procedure including gel filtration, weak cation exchange HPLC and reverse phase HPLC. All steps are carried out at low pH using exclusively volatile acidic buffer solutions. The sterile conditions and easy removal of salt by lyophilization facilitate the quantification of the growth factor in biological assays. Based on immunological characterization the purified acid-stable, highly basic transforming growth factor beta is the beta 1 form. Using the present method pure platelet TGF beta 1 is obtained in very high yield. 40 units of outdated human platelets yield 800 micrograms pure TGF beta 1, which is about a 10-20 fold higher yield than reported for other purification procedures.
已开发出一种从人血小板中纯化β型转化生长因子的新方法。该方法为三步法，包括凝胶过滤、弱阳离子交换高效液相色谱法和反相高效液相色谱法。所有步骤均在低pH值下进行，仅使用挥发性酸性缓冲溶液。无菌条件以及通过冻干易于去除盐分，这有利于在生物学测定中对生长因子进行定量。基于免疫学特征，纯化的酸稳定、高碱性的转化生长因子β为β1形式。使用本方法可获得高产率的纯血小板转化生长因子β1。40单位过期人血小板可产生800微克纯转化生长因子β1，这比其他纯化方法报道的产率高约10至20倍。
