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 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.
Department of Chemistry and Biochemistry, California State University, Los Angeles, 5151 State University Drive, Los Angeles, California 90032-8202, USA.
Mark D
MD
Goldberg
Roger C
RC
Lo
Silvija
S
Abele
Miroslav
M
Macka
Frank A
FA
Gomez
D000900:Anti-Bacterial Agents / Q000737:chemistry; D008024:Ligands; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D008856:Microscopy, Fluorescence; D009842:Oligopeptides / Q000737:chemistry*; D011437:Propylamines; D012821:Silanes / Q000737:chemistry*; D017334:Teicoplanin / Q000737:chemistry; D014640:Vancomycin / Q000737:chemistry
D000900:Anti-Bacterial Agents; D008024:Ligands; D009842:Oligopeptides; D011437:Propylamines; D012821:Silanes; C542686:carboxyfluorescein-D-Ala-D-Ala-D-Ala; D017334:Teicoplanin; D014640:Vancomycin; C477625:amino-propyl-triethoxysilane