Evaluation of the human serum albumin column as a discovery screening tool for plasma protein binding
Evaluation of the human serum albumin column as a discovery screening tool for plasma protein binding
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DOI：
10.1016/s0928-0987(01)00219-6
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发表时间：
2002-03-01
影响因子：
4.6
通讯作者：
Cohen, L
中科院分区：
文献类型：
--
作者：
Buchholz, L;Cai, CH;Cohen, L
使用人血清白蛋白柱结合UV和质谱检测，共测定了69种具有各种化学结构的化合物。中度相关性，R-2=0.661。观察了通过传统的平衡透析或超滤技术测定的血浆蛋白结合率与色谱保留因子（k '/k' + 1）之间的关系。在整个血浆蛋白结合率范围内观察到回归线和许多样本之间的差异。尝试从数据集中区分化合物以实现基于物理化学性质的更好的相关性是不成功的。采用含磷酸盐缓冲液的移动的相进行UV检测和采用含乙酸盐缓冲液的移动的相进行质谱检测所得保留时间具有良好的一致性。对于最小或高度结合（>90%结合）的化合物，对于在单态或盒中分析的化合物，获得了基本相同的数据。对含有血浆蛋白结合率大于90%的化合物的试剂卡进行分析时，即使分析物浓度高达100 μ g/ml，也不会导致色谱柱过载。不同的结果时，色谱保留值被用来排序各类化合物。当一个化合物类包含广泛的蛋白质结合时，与给定类内的化合物都高度结合相比，获得了与已知结合的排序更好的相关性。(C)2002 Elsevier Science B. V.保留所有权利。
A total of 69 compounds with a variety of chemical structures were assayed using a human serum albumin column in combination with UV and mass spectrometric detection. A moderate correlation, R-2=0.661. between the plasma protein binding, determined by traditional techniques of equilibrium dialysis or ultrafiltration, and chromatographic retention factor (k'/k' + 1) was observed. Disparity between the regression line and numerous samples was observed across the entire range of plasma protein binding. Attempts to discriminate between compounds from the data set to achieve better correlation based physico-chemical properties were unsuccessful. Good agreement was observed for retention times obtained with UV detection with mobile phase containing phosphate buffer and mass spectrometric detection with mobile phase containing acetate buffer. Essentially identical data were obtained for compounds analyzed in singlet or cassette for minimally or highly bound (>90% bound) compounds. Analysis of cassettes containing compounds with plasma protein binding greater than 90% did not cause column overload, even at analyte concentrations up to 100 mug/ml. Diverse results were obtained when chromatographic retention was used to rank order various classes of compounds. Better correlation with ordering from known binding was obtained when a compound class contained a wide range of protein binding, in contrast to when compounds within a given class were all highly bound. (C) 2002 Elsevier Science B.V. All rights reserved.