Development of SPR Biosensor Apply to the Drug Screening Process - Focus on CYP2D6 Inhibition Assay
Date Issued
2006
Date
2006
Author(s)
Lin, Jing-Rung
DOI
en-US
Abstract
The advantage of surface plasmon resonance (SPR) biosensor includes label-free and high sensitivity. Due to the characteristics of SPR biosensor and the requirement of high throughput screening (HTS) in drug development, the aim of this study is to develop SPR biosensor for rapid screening in CYP450 inhibition assay.
Compared with the common fluorescence-based method, SPR biosensor can neglect the label step in procedure and accelerate the assay speed. CYP2D6 is an important isoenzyme responsible for a large part of marked drugs. Pharmacogenetic Polymorphism is also observed in CYP2D6. Here the measurement of 50% inhibitory concentration (IC50) for CYP2D6 with non-inhibitor (dextromethorphan) and inhibitor (quinidine) using SPR biosensor is demonstrated. The probe drug, debrisoquine, is immobilized on sensor surface to detect the activity of CYP2D6. Nonspecific binding can lead to measurement error. Here the magnitude of error is also measured and the results are corrected, and the correlation between enzyme activity and binding reaction is also observed. Results of IC50 measurement are 1.3456 μM and 0.32129 nM for dextromethorphan and quinidine. After correction, IC50 of dextromethorphan and quinidine are 0.8624 μM and 0.07814 nM, both are smaller than uncorrected value. It shows that testing substrate concentration is associated with binding reaction and enzyme activity. This study represents that SPR biosensor can be considered as a tool for CYP450 inhibition assay after the selected probe drugs are immobilized on sensor surface. These results show the difference between the non-inhibitor and the inhibitior. From the results of IC50, the distinction between non-inhibitor and inhibitor are observed. The unavoidable error can be estimated and used for correcting IC50 measurement results.
Because the multi-channel design and automation can be carried out in SPR biosensor, the results of this study suggest that biosensor based on SPR has the potential to provide a HTS tool for drug screening in the future.
Subjects
表面電漿共振生物感測器
CYP450
50%抑制濃度
SPR biosensor
IC50
Type
thesis
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