Baier G, Wollensak G, Mur E, Redl B, Stöffler G, Göttinger W
Institut für Mikrobiologie, Medizinischen Fakultät, Universität Innsbruck, Austria.
J Chromatogr. 1990 Feb 23;525(2):319-28. doi: 10.1016/s0378-4347(00)83408-8.
A comparison of the efficiencies of hydrophobic interaction chromatography, ion-exchange chromatography, reversed-phase chromatography and gel permeation chromatography in the separation of tear proteins was made using a variety of different buffers. Separation of immunoglobulins, lactoferrin, albumin, PMFA (protein migrating faster than albumin) and lysozyme was accomplished by gel permeation chromatography in less than 30 min using a TSK-type SW3000 column equilibrated with ammonium acetate buffer (pH 4.1) with a high reproducibility. When gel permeation chromatography was used as a completely automated diagnostic method, only minute volumes (1.0 microliter) of tear samples were necessary for the quantitative analysis of proteins. The other three methods proved to be more suitable for the preparation of individual tear proteins but were less suitable for their quantitation.
使用多种不同缓冲液，对疏水相互作用色谱法、离子交换色谱法、反相色谱法和凝胶渗透色谱法分离泪液蛋白质的效率进行了比较。使用用醋酸铵缓冲液（pH 4.1）平衡的TSK型SW3000柱，通过凝胶渗透色谱法在不到30分钟内完成了免疫球蛋白、乳铁蛋白、白蛋白、PMFA（迁移速度比白蛋白快的蛋白质）和溶菌酶的分离，重现性高。当凝胶渗透色谱法用作完全自动化诊断方法时，仅需微量体积（1.0微升）的泪液样本即可进行蛋白质的定量分析。事实证明，其他三种方法更适合于制备单个泪液蛋白质，但不太适合其定量分析。
