title: Fast measurement of binding kinetics with dual slope SPR microchips.
abstract: We demonstrate a new dual slope SPR technique that is ten-fold faster than the conventional step-response method. The new scheme utilizes rapid slope-based measurements followed by rapid reset, and it separates association and dissociation half reaction measurements at two separate sites inside a dual-chamber PDMS microfluidic chip. For a model CAII-ABS test system, the association and dissociation slopes were measured in 30 seconds compared to 5 minutes for step-response. The values of k(a) and k(d) calculated from the slope method are 3.66 ± 0.19 × 10(3) M(-1) s(-1) and 4.83 ± 0.17 × 10(-2) s(-1), respectively, matching well with step-response values while facilitating ~10 to 15 fold faster detection and quantification.
authors.affiliation: Department of Bioengineering, University of Utah, Salt Lake City, UT, USA. t.ghosh@utah.edu
authors.forename: Tridib
authors.initials: T
authors.lastname: Ghosh
authors.forename: Carlos H
authors.initials: CH
authors.lastname: Mastrangelo
mesh_terms: D024402:Carbonic Anhydrase II / Q000378:metabolism; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D007700:Kinetics; D046228:Microarray Analysis; D046210:Microfluidic Analytical Techniques; D059629:Signal-To-Noise Ratio; D020349:Surface Plasmon Resonance
chemical_list: D004129:Dimethylpolysiloxanes; C013830:baysilon; D024402:Carbonic Anhydrase II