Fast measurement of binding kinetics with dual slope SPR microchips

Analyst. 2012 May 21;137(10):2381-5. doi: 10.1039/c2an35045a. Epub 2012 Apr 11.

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.

MeSH terms

  • Carbonic Anhydrase II / metabolism
  • Dimethylpolysiloxanes / chemistry
  • Kinetics
  • Microarray Analysis*
  • Microfluidic Analytical Techniques
  • Signal-To-Noise Ratio
  • Surface Plasmon Resonance*

Substances

  • Dimethylpolysiloxanes
  • baysilon
  • Carbonic Anhydrase II