Analysis of drug-protein binding by ultrafast affinity chromatography using immobilized human serum albumin.
Analysis of drug-protein binding by ultrafast affinity chromatography using immobilized human serum albumin.
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DOI：
10.1016/j.chroma.2010.02.026
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发表时间：
2010-04-23
期刊：
影响因子：
--
通讯作者：
Hage DS
中科院分区：
文献类型：
--
作者：
Mallik R;Yoo MJ;Briscoe CJ;Hage DS
人血清白蛋白（HSA）被探索用作固定相和配体的亲和微柱的超快提取游离药物馏分和使用此信息的药物-蛋白结合的分析。在本研究中，华法林、丙咪嗪和布洛芬被用作模型分析物。结果发现，大于95%的所有这些药物的提取可以实现在短短的250 ms的HSA微柱。保留的药物馏分，然后从相同的柱在等度条件下洗脱，在4.5 mL/min的工作时，在不到40 s的洗脱。该系统的色谱行为给出了一个很好的拟合与计算机模拟的基础上预测的可逆的，饱和模型的注射药物与固定化HSA的结合。通过这种方法测得的游离馏分被认为是可比的超滤测定，并通过这种方法估计的平衡常数与文献值有很好的一致性。该方法的优点包括其速度和含有HSA的微柱相对较低的成本。HSA结合多种类型药物的能力也创造了使用相同的亲和微柱来研究和测量各种药剂的游离级分的可能性。这些特性使得该技术在药物结合研究和新药候选物的高通量筛选中具有吸引力。
Human serum albumin (HSA) was explored for use as a stationary phase and ligand in affinity microcolumns for the ultrafast extraction of free drug fractions and the use of this information for the analysis of drug-protein binding. Warfarin, imipramine, and ibuprofen were used as model analytes in this study. It was found that greater than 95% extraction of all these drugs could be achieved in as little as 250 ms on HSA microcolumns. The retained drug fraction was then eluted from the same column under isocratic conditions, giving elution in less than 40 s when working at 4.5 mL/min. The chromatographic behavior of this system gave a good fit with that predicted by computer simulations based on a reversible, saturable model for the binding of an injected drug with immobilized HSA. The free fractions measured by this method were found to be comparable to those determined by ultrafiltration, and equilibrium constants estimated by this approach gave good agreement with literature values. Advantages of this method include its speed and the relatively low cost of microcolumns that contain HSA. The ability of HSA to bind many types of drugs also creates the possibility of using the same affinity microcolumn to study and measure the free fractions for a variety of pharmaceutical agents. These properties make this technique appealing for use in drug binding studies and in the high-throughput screening of new drug candidates.
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影响因子：
3.1
作者：
Hage, David S.;Jackson, Abby;Sobansky, Matthew R.;Schiel, John E.;Yoo, Michelle J.;Joseph, K. S.
通讯作者：
Joseph, K. S.
影响因子：
2.1
作者：
Kuroda, Yukihiro;Saito, Madoka;Yamaoka, Takashi
通讯作者：
Yamaoka, Takashi
影响因子：
7.4
作者：
Hage, DS;Nelson, MA
通讯作者：
Nelson, MA
影响因子：
3.8
作者：
Day, YSN;Myszka, DG
通讯作者：
Myszka, DG
影响因子：
4.1
作者：
HAGE, DS;NOCTOR, TAG;WAINER, IW
通讯作者：
WAINER, IW