Real-time observation of affinity reactions using grating couplers: determination of the detection limit and calculation of kinetic rate constants

Analytical Biochemistry
R PolziusU Bilitewski

Abstract

The use of integrated optical grating couplers for the analysis of bioaffinity reactions in order to calculate kinetic rate constants was investigated. The specificity of the sensor surface was determined by adsorptive or covalent attachment of the specific ligands. As an evanescent field sensor, the specific interaction of the corresponding ligand could be observed in real time and without labels. The detection limit in terms of the molecular weight of the analyte was studied by the specific binding of biotinylated proteins of different molecular weights to avidin-loaded sensors. It was shown that grating coupler sensors allowed detection of compounds of at least 2000 daltons using high-affinity receptors, while the direct sensing of low molecular analytes, such as biotin, could not be significantly achieved. Association rate constants were calculated for the interaction of the different biotinylated proteins to avidin-covered sensors from single binding curves. Due to the strong binding between avidin and biotin, the dissociation of the formed complex could not be observed. Kinetic rate constants and equilibrium constants were determined by studying the interaction of human immunoglobulin with the immobilized receptor, protei...Continue Reading

References

Jan 1, 1991·Biosensors & Bioelectronics·W LukoszP Weiss
Oct 4, 1988·Journal of Immunological Methods·M Stenberg, H Nygren
Jan 1, 1996·Biosensors & Bioelectronics·R PolziusW Koschinski
Dec 1, 1993·Biotechnology and Bioengineering·R PolziusR D Schmid

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Citations

Apr 18, 2007·Analytical Chemistry·Jennifer O FoleyPaul Yager
Mar 25, 2005·Artificial Organs·Isa Santos DuarteSonia Maria Alves Bueno
Jun 7, 2008·Biosensors & Bioelectronics·Namsoo KimWoo-Yeon Kim
Jan 5, 2006·Optics Letters·A Ksendzov, Y Lin
Jan 6, 2008·Sensors·Katrin SchmittChristian Hoffmann
Jul 1, 2011·Optics Express·Xing Wei, Sharon M Weiss
Nov 24, 2001·Clinica Chimica Acta; International Journal of Clinical Chemistry·P B LuppaD W Chan
Jun 8, 2002·Biosensors & Bioelectronics·A Akkoyun, U Bilitewski
Mar 6, 1999·Journal of Chromatography. B, Biomedical Sciences and Applications·J Turková
May 3, 2006·Langmuir : the ACS Journal of Surfaces and Colloids·John R LaGraff, Quynh Chu-LaGraff
Jun 15, 2011·Biosensors & Bioelectronics·Konstantinos Mitsakakis, Electra Gizeli

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