Noncompetitive peak decay analysis of drug-protein dissociation by high-performance affinity chromatography.
Noncompetitive peak decay analysis of drug-protein dissociation by high-performance affinity chromatography.
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DOI：
10.1002/jssc.200900074
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发表时间：
2009-05
影响因子：
3.1
通讯作者：
Hage, David S.
中科院分区：
文献类型：
--
作者：
Chen, Jianzhong;Schiel, John E.;Hage, David S.
关键词：
峰衰减法是一种亲和色谱技术，已用于检查在过量置换剂存在下溶质从固定化配体上解离的情况。然而，可能难以找到不干扰洗脱分析物的检测的置换剂。在这项研究中，开发了一种非竞争性峰衰减方法，其中不需要置换剂用于分析物洗脱。该方法被评价为药物-蛋白质相互作用的研究，通过使用它沿着与高效亲和色谱法，以测量解离速率常数的R-和S-华法林从含有固定化人血清白蛋白（HSA）的柱。在该方法的优化中考虑了几个因素，包括所应用的分析物的量、柱尺寸和流速。通过这种方法在几个温度下测量了R-和S-华法林从HSA的解离速率常数，在pH 7.4和37°C下的值分别为0.56（± 0.01）和0.66（± 0.01）s−1。这些结果与以前的其他方法得到的值吻合得很好。这种方法不仅限于华法林和HSA，还可以用于研究其他药物-蛋白质相互作用或其他具有弱至中度结合的系统。
The peak decay method is an affinity chromatographic technique that has been used to examine the dissociation of solutes from immobilized ligands in the presence of excess displacing agent. However, it can be difficult to find a displacing agent that does not interfere with detection of the eluting analyte. In this study, a non-competitive peak decay method was developed in which no displacing agent was required for analyte elution. This method was evaluated for the study of drug-protein interactions by using it along with high-performance affinity chromatography to measure the dissociation rate constants for R- and S-warfarin from columns containing immobilized human serum albumin (HSA). Several factors were considered in the optimization of this method, including the amount of applied analyte, the column size, and the flow rate. The dissociation rate constants for R- and S-warfarin from HSA were measured at several temperatures by this approach, giving values of 0.56 (± 0.01) and 0.66 (± 0.01) s−1 at pH 7.4 and 37°C. These results were in good agreement with previous values obtained by other methods. This approach is not limited to warfarin and HSA but could be employed in studying additional drug-protein interactions or other systems with weak-to-moderate binding.
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影响因子：
4.1
作者：
RUHN, PF;GARVER, S;HAGE, DS
通讯作者：
HAGE, DS
影响因子：
5.8
作者：
BOS, OJM;FISCHER, MJE;JANSSEN, LHM
通讯作者：
JANSSEN, LHM
影响因子：
4.1
作者：
Chattopadhyay, A;Hage, DS
通讯作者：
Hage, DS
影响因子：
3.6
作者：
RIETBROCK, N;LASSMANN, A
通讯作者：
LASSMANN, A
DOI：
10.1016/s0021-9673(01)87800-6
发表时间：
1984-01-01
期刊：
JOURNAL OF CHROMATOGRAPHY
影响因子：
--
作者：
MULLER, AJ;CARR, PW
通讯作者：
CARR, PW