Fast measurement of binding kinetics with dual slope SPR microchips |
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Authors: | Ghosh Tridib Mastrangelo Carlos H |
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Institution: | Department of Bioengineering, University of Utah, Salt Lake City, UT, USA. t.ghosh@utah.edu |
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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. |
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