化学
色谱法
洗脱
离子色谱法
分馏
离子交换
梯度洗脱
滴定曲线
滴定法
固定化pH梯度
圆周率
分析化学(期刊)
离子
高效液相色谱法
等电聚焦
生物化学
无机化学
酶
有机化学
作者
Tangir Ahamed,Beckley K. Nfor,Peter Verhaert,Gijs W. K. van Dedem,Luuk A. M. van der Wielen,Michel H. M. Eppink,Emile J.A.X. van de Sandt,Marcel Ottens
标识
DOI:10.1016/j.chroma.2007.07.010
摘要
This work demonstrates that a highly linear, controllable and wide-ranged pH-gradient can be generated through an ion-exchange chromatography (IEC) column. Such a pH-gradient anion-exchange chromatography was evaluated with 17 model proteins and found that acidic (pI < 6) and basic (pI > 8) proteins elute roughly at their pI, whereas neutral proteins (pI 6–8) elute at pH 8–9 regardless their pI values. Because of the flat nature of protein titration curves from pH ∼6 to ∼9, neutral proteins indeed exhibit nearly zero net charge at pH ∼9. The elution-pH in pH-gradient IEC or the titration curve, but not the pI, was identified as the key parameter for pH optimization of preparative IEC in a fast and rational way. The pH-gradient IEC was also applied and found to be an excellent analytical tool for the fractionation of crude protein mixtures.
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