Refractive index-based detection of biomolecular interactions

Christine Campagnolo; Thomas Ryan; Robert C. Atkinson; Carl A. Batt

doi:10.1117/12.418735

13 March 2001 Refractive index-based detection of biomolecular interactions

Christine Campagnolo, Thomas Ryan, Robert C. Atkinson, Carl A. Batt

Proceedings Volume 4206, Photonic Detection and Intervention Technologies for Safe Food; (2001) https://doi.org/10.1117/12.418735
Event: Environmental and Industrial Sensing, 2000, Boston, MA, United States

Abstract

A biosensor was constructed by modifying the optics of a critical angle refractometer to enable it to measure changes in surface plasmon resonance. Initially, the utility of the critical angle refractometry to measure biomolecular interactions was examined using the Leica Automatic Critical Angle Refractometer 600 (AR600). While initial results suggested that the biomolecular interactions could be measured by critical angle refractometry, these proved to be an artifact of protein buildup on the surface. At lower concentrations of target protein, no change in the refractive index was observed. The AR600 was reconfigured from a critical angle refractometer to surface plasmon resonance technology to study molecular interactions between biomolecules. The performance of the SPR biosensor was demonstrated by measuring protein-protein interactions; specifically, avidin:anti-avidin interactions and streptavidin:anti-streptavidin interactions. Equilibrium dissociation constants were obtained by data analysis according to pseudo-first order binding kinetics. Polyclonal anti-streptavidin and monoclonal antiavidin demonstrated affinity constants of 9.7 x 10-5 M-1 and 4.3 x 10-5 M-1 for their respective ligands.

Citation Download Citation

Christine Campagnolo, Thomas Ryan, Robert C. Atkinson, and Carl A. Batt "Refractive index-based detection of biomolecular interactions", Proc. SPIE 4206, Photonic Detection and Intervention Technologies for Safe Food, (13 March 2001); https://doi.org/10.1117/12.418735

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