Development of microfluidic chips for heterogeneous receptor-ligand interaction studies

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 × 10 ⁴ and 4.93 ± 1.13 × 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.