Evaluation of microcolumn stability in ultrafast affinity extraction for binding and rate studies.
Evaluation of microcolumn stability in ultrafast affinity extraction for binding and rate studies.
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用于结合和速率研究的超快亲和萃取中微柱稳定性的评价。
DOI：
10.1016/j.jchromb.2021.123047
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发表时间：
2021-12-15
期刊：
影响因子：
--
通讯作者：
Hage DS
中科院分区：
文献类型：
--
作者：
Iftekhar S;Hage DS
超快亲和萃取（UAE）最近被开发并用于测量药物和其他可溶结合剂（如血清蛋白）的非结合（或游离）部分和结合或速率常数。本研究检测了10 mm × 2.1 mm亲和微柱在低流速和高流速（例如，0.5和3.5 mL/min）下用于UAE时的长期稳定性。以固定的人血清白蛋白（HSA）和含华法林（含或不含可溶性HSA）的样品为模型系统，研究了这种稳定性。在可溶性HSA存在下，以0.5 mL/min测量的游离华法林组分在注射150针时稳定，在3.5 mL/min时变化< 10%。阿联酋估计华法林与HSA的关联平衡常数为3.5 mL/min，在50次注射中没有显著变化，在100-150次注射中变化仅为~ 18-22%。结合0.5 mL/min和3.5 mL/min的UAE结果，并采用新的两点法，发现华法林与HSA的解离速率常数，即使在200次注射后，该值也没有明显变化。延长微柱的使用对华法林的保留时间、峰宽、峰不对称以及微柱背压的影响也进行了考虑。这些结果表明，阿联酋和HSA微柱可以在大范围的注射中提供一致的自由溶质分数、结合常数和速率常数。这些结果将有助于未来的工作，为评估、进一步开发和使用UAE来表征溶液样品中涉及其他药物和结合剂的相互作用提供指导。
Ultrafast affinity extraction (UAE) has recently been developed and employed for measuring non-bound (or free) fractions and binding or rate constants for drugs and other targets with soluble binding agents such as serum proteins. This study examined the long-term stability of 10 mm × 2.1 mm i.d. affinity microcolumns when used in UAE at both low and high flow rates (e.g., 0.5 and 3.5 mL/min) over an extended series of injections. This stability was investigated by using immobilized human serum albumin (HSA) and samples containing the drug warfarin with or without soluble HSA as a model system. The free warfarin fractions measured at 0.5 mL/min in the presence of soluble HSA were stable up to 150 injections and changed by < 10% at 3.5 mL/min. The association equilibrium constant for warfarin with HSA that was estimated by UAE at 3.5 mL/min had no significant change over 50 injections and a change of up to only ~18–22% over 100–150 injections. The dissociation rate constant for warfarin from HSA was found by combining UAE results at 0.5 and 3.5 mL/min and employing a new two-point approach, with no significant changes in this value being seen even after 200 injections. The effects of extended microcolumn use on the retention time, peak width, and peak asymmetry for warfarin, and on the backpressure of the microcolumn, were also considered. These results indicated that UAE and HSA microcolumns could be used to provide consistent values for free solute fractions, binding constants, and rate constants over a large series of injections. These results should be useful in future work by providing guidelines for the assessment, further development, and use of UAE in characterizing interactions involving other drugs and binding agents in solution-based samples.
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影响因子：
4.1
作者：
Kirkland, JJ;Henderson, JW;Claessens, HA
通讯作者：
Claessens, HA
DOI：
10.1016/j.chroma.2010.02.026
发表时间：
2010-04-23
期刊：
Journal of chromatography. A
影响因子：
--
作者：
Mallik R;Yoo MJ;Briscoe CJ;Hage DS
通讯作者：
Hage DS
影响因子：
7.4
作者：
FOLEY, JP;DORSEY, JG
通讯作者：
DORSEY, JG
影响因子：
8.4
作者：
Czub, Mateusz P.;Venkataramany, Barat S.;Minor, Wladek
通讯作者：
Minor, Wladek
DOI：
10.1016/j.chroma.2017.03.056
发表时间：
2017-05-12
期刊：
Journal of chromatography. A
影响因子：
--
作者：
Beeram S;Bi C;Zheng X;Hage DS
通讯作者：
Hage DS