Paquet A, Lévesque A, Pagé M
Department of Biochemistry, Faculty of Medicine, Université Laval, Sainte-Foy, Québec, Canada.
J Chromatogr A. 1994 Apr 29;667(1-2):125-30. doi: 10.1016/0021-9673(94)89060-9.
Human recombinant tumor necrosis factor alpha was purified in a single step to about 95% purity from Escherichia coli lysate by chromatography on hydroxyapatite. The last traces of contaminants were removed by fast protein liquid chromatography on a Mono Q column. The final product was found to be pure by gel electrophoresis with silver staining. A molecular mass of approximately 17,000 and a specific activity of 4.3 x 10(6) U/mg after a single purification step were found.
通过羟基磷灰石色谱法从大肠杆菌裂解物中一步纯化出纯度约为95%的重组人肿瘤坏死因子α。最后痕量的污染物通过在Mono Q柱上进行快速蛋白质液相色谱法去除。通过银染凝胶电泳发现最终产物是纯的。单次纯化步骤后发现其分子量约为17,000,比活性为4.3×10⁶U/mg。