Purification of transforming growth factor type e.
Purification of transforming growth factor type e.
复制标题
e型转化生长因子的纯化。
DOI：
10.1002/jcb.240420207
复制
发表时间：
1990
影响因子：
4
通讯作者：
Halper,J
中科院分区：
文献类型：
--
作者：
Parnell,PG;Wunderlich,J;Carter,B;Halper,J
e 型转化生长因子 (TGFe) 是一种热和酸稳定的多肽，表观分子量为 22,000，可刺激单层和软琼脂中某些上皮和间质细胞的增殖。 TGFe 已纯化至均质。对牛肾进行初始酸乙醇提取，然后使用 Bio Rex 70 树脂进行批量离子交换色谱。使用配备SIY10螺旋膜的Amicon浓缩器对从Bio Rex 70树脂洗脱的活性进行浓缩和渗滤，然后通过Bio-Gel P-60分子筛色谱进一步纯化。收集分子筛色谱的活性组分，并通过肝素-琼脂糖亲和色谱纯化，然后使用微孔 C-8 柱进行反相高效液相色谱。最后的纯化步骤涉及通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分离的 TGFe 的电洗脱。 TGFe 的纯度经评估大于 90%。
Transforming growth factor type e (TGFe) is a heat‐ and acid‐stable polypeptide with an apparent molecular weight of 22,000, which stimulates the proliferation of certain epithelial and mesenchymal cells in monolayer and soft agar. TGFe has been purified to homogeneity. Initial acid‐ethanol extraction of bovine kidney was followed by batch ion‐exchange chromatography utilizing Bio Rex 70 resin. The activity eluted from the Bio Rex 70 resin was concentrated and diafiltered using an Amicon concentrator equipped with an SIY10 spiral membrane, then was further purified by Bio‐Gel P‐60 molecular sieve chromatography. Active fractions from molecular sieve chromatography were pooled and purified by heparin‐Sepharose affinity chromatography, followed by reverse‐phase high‐performance liquid chromatography using a microbore C‐8 column. The final purification step involved electroelution of TGFe separated by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE). Purity of TGFe was assessed to be greater than 90%.