Improved protein recovery in reversed-phase liquid chromatography by the use of ultrahigh pressures
Improved protein recovery in reversed-phase liquid chromatography by the use of ultrahigh pressures
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DOI：
10.1021/ac0518304
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发表时间：
2006-03-01
影响因子：
7.4
通讯作者：
Jorgenson, JW
中科院分区：
文献类型：
--
作者：
Eschelbach, JW;Jorgenson, JW
研究了超高压液相色谱（UHPLC）中压力升高对蛋白质回收率的影响。具体地，检查蛋白质残留（“重影”）和回收率。在常规压力（160 bar）和超高压（> 1500 bar）下，通过梯度RPLC分离四种模型蛋白质（核糖核酸酶A、卵清蛋白、肌红蛋白、BSA）。一个定制的梯度UHPLC系统被用来产生常规的压力5 μ m直径的反相支持和相同的1.4 μ m的支持上的反相压力。结果表明，通过增加压力，运行之间的蛋白质携带减少，并且在某些情况下消除了所研究的模型蛋白质的特定阈值压力以上。进一步的研究表明，每种研究的蛋白质的回收率都得到了提高，某些蛋白质的回收率甚至接近100%。
The effect that elevated pressure used in ultrahigh-pressure liquid chromatography (UHPLC) has on protein recovery was investigated. Specifically, protein carryover ("ghosting") and recovery were examined. Four model proteins (ribonuclease A, ovalbumin, myoglobin, BSA) were separated by gradient RPLC at both conventional (160 bar) and ultrahigh pressures (> 1500 bar). A custom gradient UHPLC system was used to generate conventional pressures on 5-mu m diameter reversed-phase supports and ultrahigh pressures on identical 1.4-mu m supports. The results indicate that, by increasing the pressure, protein carryover from run to run is reduced and in some cases eliminated above a certain threshold pressure for the model proteins studied. Further work indicates that recovery was enhanced for each of the proteins studied, even approaching 100% for certain proteins.