Real-Time Biomolecular Binding Detection Using a Sensitive Photonic Crystal Biosensor

Real-time measurement of specific biomolecular interactions is critical to many areas of biological research. A number of label-free techniques for directly monitoring biomolecular binding have been developed, but it is still challenging to measure the binding kinetics of very small molecules, to de...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2010-06, Vol.82 (12), p.5211-5218
Main Authors: Guo, Yunbo, Ye, Jing Yong, Divin, Charles, Huang, Baohua, Thomas, Thommey P, Baker, Jr, James R, Norris, Theodore B
Format: Article
Language:English
Subjects:
Analytical chemistry
Antibodies - metabolism
Binding Sites
Biosensing Techniques - methods
Biosensors
Biotin - metabolism
Biotinylation
Molecular chemistry
Molecular weight
Molecules
Oligonucleotides - metabolism
Photons
Protein Binding
Proteins
Proteins - metabolism
Sensitivity and Specificity
Streptavidin - metabolism
Studies
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Summary:Real-time measurement of specific biomolecular interactions is critical to many areas of biological research. A number of label-free techniques for directly monitoring biomolecular binding have been developed, but it is still challenging to measure the binding kinetics of very small molecules, to detect low concentrations of analyte molecules, or to detect low affinity interactions. In this study, we report the development of a highly sensitive photonic crystal biosensor for label-free, real-time biomolecular binding analysis. We characterize the performance of this biosensor using a standard streptavidin−biotin binding system. Optimization of the surface functionalization methods for streptavidin immobilization on the silica sensing surface is presented, and the specific binding of biotinylated analyte molecules ranging over 3 orders of magnitude in molecular weight, including very small molecules (150 000 Da), are detected in real time with a high signal-to-noise ratio. Finally, we document the sensor efficiency for low mass adsorption, as well as multilayered molecular interactions. By all important metrics for sensitivity, we anticipate this photonic crystal biosensor will provide new capabilities for highly sensitive measurements of biomolecular binding.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac100576y