Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
7559694 | Analytical Biochemistry | 2012 | 6 Pages |
Abstract
The measurement of membrane affinity is an important early screening step during drug discovery. However, classical methods for membrane affinity measurement are tedious and difficult to implement in high-throughput screening. This article describes a quantitative method for the measurement of membrane affinity by colorimetric assay based on polydiacetylene (PDA) sensors. Prepared lipid/PDA chromatic vesicles were used to model cell membranes. By measuring the colorimetric response of the chromatic vesicles when drug-membrane interactions occurred, membrane affinity constant Kb could be calculated using a simple quantitative model. Under optimized preparation conditions, the calculated log(Kb) values exhibited an in-batch relative standard deviation (RSD) of less than 4% and a between-batch RSD of less than 8% for all three reference compounds. The logarithm of Kb of the six β-blockers exhibited excellent linear correlation with the logarithm of the liposome/water partition coefficient (Km) with R2 = 0.9793. For neutral compounds, the log(Kb) of n-fatty alcohols correlated with the logarithm of the n-octanol/water partition coefficient (Koct) with a linear correlation coefficient R2 = 0.9833. This work provides a simple, convenient, and reproducible method for the rapid measurement of membrane affinity and presents important implications for high-throughput screening.
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Feng Zheng, Zheng Wu, Yihua Chen,