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Anal Chem. 2009 Jun 15;81(12):5095-8. doi: 10.1021/ac9006649.

Development of microfluidic chips for heterogeneous receptor-ligand interaction studies.

Author information

1
Department of Chemistry and Biochemistry, California State University, Los Angeles, 5151 State University Drive, Los Angeles, California 90032-8202, USA.

Abstract

A simple microfluidic-based technique to quantitate the binding affinity between the glycopeptide antibiotics teicoplanin from Actinoplanes teicomyceticus and vancomycin from Streptomyces orientalis and 5-carboxyfluorescein-D-Ala-D-Ala-D-Ala (5-FAM-(DA)(3)) is described. In this work, (3-aminopropyl)triethoxysilane is used to modify the surfaces of a series of microchannels, and each channel is subsequently exposed to a solution of antibiotic for a few minutes. The antibiotic is retained after washing through electrostatic interactions, and the series of channels are subsequently exposed to an increasing concentration of 5-FAM-(DA)(3) followed by washing to exclude any nonspecific binding. The extent of fluorescence is quantified using a microscope fitted with a CCD camera. The binding constants for the interaction of teicoplanin and vancomycin with the fluorescent peptide were determined to be 6.03 +/- 0.97 x 10(4) and 4.93 +/- 1.13 x 10(4) M(-1), respectively, in good agreement with previous data. The ease of quantifying the extent of interaction in this microchip technique may prove powerful for exploration of a myriad of receptor-ligand pairs.

PMID:
19441833
DOI:
10.1021/ac9006649
[Indexed for MEDLINE]

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