Please use this identifier to cite or link to this item: https://doi.org/10.1016/j.jchromb.2005.01.015
Title: Development and characterization of an immobilized human organic cation transporter based liquid chromatographic stationary phase
Authors: Moaddel, R.
Yamaguchi, R.
Ho, P.C. 
Patel, S.
Hsu, C.-P.
Subrahmanyam, V.
Wainer, I.W.
Keywords: Affinity chromatography
Drug transporters
hOCT1
MDCK cells
Issue Date: 25-Apr-2005
Source: Moaddel, R., Yamaguchi, R., Ho, P.C., Patel, S., Hsu, C.-P., Subrahmanyam, V., Wainer, I.W. (2005-04-25). Development and characterization of an immobilized human organic cation transporter based liquid chromatographic stationary phase. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences 818 (2) : 263-268. ScholarBank@NUS Repository. https://doi.org/10.1016/j.jchromb.2005.01.015
Abstract: Membranes from a stably transfected cell line that expresses the human organic cation 1 transporter (hOCT1) have been immobilized on the immobilized artificial membrane (IAM) liquid chromatographic stationary phase to form the hOCT1(+)-IAM stationary phase. Membranes from the parent cell line that does not express the hOCT1 were also immobilized to create the hOCT1(-)-IAM stationary phase. Columns were created using both stationary phases, and frontal displacement chromatography experiments were conducted using [ 3H]-methyl phenyl pyridinium ([3H]-MPP+) as the marker ligand and MPP+, verapamil, quinidine, quinine, nicotine, dopamine and vinblastin as the displacers. The Kd values calculated from the chromatographic studies correlated with previously reported K i values (r2 = 0.9987; p < 0.001). The data indicate that the hOCT1(+)-IAM column can be used for the on-line determination of binding affinities to the hOCT1 and that these affinities are comparable to those obtained using cellular uptake studies. In addition, the chromatographic method was able to identify a previously undetected high affinity binding site for MPP+ and to determine that hOCT1 bound (R)-verapamil to a greater extent than (S)-verapamil.
Source Title: Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
URI: http://scholarbank.nus.edu.sg/handle/10635/105816
ISSN: 15700232
DOI: 10.1016/j.jchromb.2005.01.015
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