Lihme A, Heegaard P M
Research Center for Medical Biotechnology, University of Copenhagen, Denmark.
Anal Biochem. 1991 Jan;192(1):64-9. doi: 10.1016/0003-2697(91)90184-u.
Human serum proteins were separated on a matrix obtained by reaction of beta-mercaptoethanol with divinyl sulfone-activated agarose (the so-called T-gel). Binding of almost all serum proteins was observed at high concentrations of ammonium sulfate. Elution was achieved by gradually lowering the concentration of salt in the washing buffer. Fractions obtained during elution were analyzed by fused rocket immunoelectrophoresis. Proteins were recovered in high yields and with an excellent separation in this one-step procedure ("Thiophilic adsorption chromatography"). A rapid and straightforward procedure giving essentially pure immunoglobulins from crude rabbit serum with at least 80% yield by the T-gel is also presented.
人血清蛋白在由β-巯基乙醇与二乙烯砜活化琼脂糖反应得到的基质(即所谓的T凝胶)上进行分离。在高浓度硫酸铵条件下观察到几乎所有血清蛋白的结合。通过逐渐降低洗涤缓冲液中盐的浓度实现洗脱。洗脱过程中得到的级分通过融合火箭免疫电泳进行分析。在这个一步法程序(“嗜硫吸附色谱法”)中,蛋白质以高收率回收且分离效果极佳。还介绍了一种快速简便的方法,通过T凝胶从粗兔血清中获得基本纯的免疫球蛋白,收率至少为80%。