Wu S L, Hancock W S, Pavlu B, Gellerfors P
Medicinal and Analytical Chemistry Department, Genentech, Inc., South San Francisco, CA 94080.
J Chromatogr. 1990 Feb 2;500:595-606. doi: 10.1016/s0021-9673(00)96094-1.
This publication analyzes different preparations of recombinant human growth hormone (rhGH) by hydrophobic-interaction chromatography (HIC). The effect of temperature on the separation was investigated as well as a series of commercially available HIC columns (TSK-phenyl-5PW, TSK-ether-PW, Beckman CAA-HIC and polypropyl A). The TSK-ether column gave the best results in the analysis of rhGH samples at different temperatures, as well as allowing an efficient separation of methionyl-hGH from rhGH. The TSK-phenyl column can be effectively used in the examination of different human growth hormone variants. The details of sample preparation have been demonstrated to be important in HIC analysis of hGH on the TSK-ether-5PW column. Injection volume and the solvent used to dissolve the protein sample are both crucial factors in this analysis. Also protein aggregation may play a role in these observations. The effect of temperature, protein concentration and spectroscopic data on the eluted protein suggest, however, that aggregation is not the cause of frontal peaks.
本出版物通过疏水相互作用色谱法（HIC）分析了重组人生长激素（rhGH）的不同制剂。研究了温度对分离的影响以及一系列市售的HIC柱（TSK-苯基-5PW、TSK-醚-PW、贝克曼CAA-HIC和聚丙烯A）。TSK-醚柱在不同温度下对rhGH样品的分析中给出了最佳结果，并且能够有效地从rhGH中分离出甲硫氨酰-hGH。TSK-苯基柱可有效地用于检测不同的人生长激素变体。已证明样品制备的细节在TSK-醚-5PW柱上hGH的HIC分析中很重要。进样体积和用于溶解蛋白质样品的溶剂都是该分析中的关键因素。此外，蛋白质聚集可能在这些观察结果中起作用。然而，温度、蛋白质浓度和光谱数据对洗脱蛋白质的影响表明，聚集不是前沿峰的原因。
