Said H M, Newsom A E, Tippins B L, Mathews R A
J Chromatogr. 1985 May 17;324(1):65-73. doi: 10.1016/s0021-9673(01)81308-x.
A new procedure for the fractionation of the heterogeneous cystine-rich proteins from human hair, utilizing reversed-phase high-performance liquid chromatography, is described. Of these proteins 27 fractions have been collected and analyzed for amino acid composition. There seems to be little correlation between the elution order and the hydrophobicity of the fraction constituents except for the late-eluting fractions. Based on the elution profiles and amino acid contents, these fractions appear to fall into four families. The effects of alkyl chain length, flow-rate and gradient slope, as well as various additives to the organic modifier on the separation have also been investigated. A low flow-rate (0.4 ml/min) and a shallow gradient were essential for the separation of these proteins as was the use of short alkyl chain (C4) or medium alkyl chain (C8) columns. However, with the C4 column reproducibility and recovery were excellent.
本文描述了一种利用反相高效液相色谱法从人发中分离富含异质胱氨酸蛋白质的新方法。已收集了这些蛋白质的27个馏分，并对其氨基酸组成进行了分析。除了洗脱较晚的馏分之外，馏分成分的洗脱顺序与疏水性之间似乎几乎没有相关性。根据洗脱曲线和氨基酸含量，这些馏分似乎可分为四类。还研究了烷基链长度、流速和梯度斜率以及向有机改性剂中添加各种添加剂对分离的影响。低流速（0.4毫升/分钟）和浅梯度对于这些蛋白质的分离至关重要，使用短烷基链（C4）或中烷基链（C8）柱也是如此。然而，使用C4柱时，重现性和回收率都非常好。
