Real-time small molecule binding detection using a label-free photonic crystal biosensor

We have demonstrated real-time, label-free detection of small molecule binding using a novel optical biosensor. This sensor is a recently developed sensing platform incorporating a one-dimensional photonic crystal (PC) structure in a total-internal-reflection (TIR) geometry (PC-TIR). This simple configuration functions as an open Fabry-Perot resonator which provides a narrow optical resonance to enable label-free, highly sensitive detection of analyte molecules on the sensing surface in the enhanced evanescent field. Moreover, when the differential intensity modulation during binding is measured, a very high detection sensitivity can be obtained, and real-time binding observed. The well-studied biotinstreptavidin system was chosen to calibrate the detection limit for small molecule detection. Effective surface functionalization methods for streptavidin immobilization on the silica sensing surface were investigated, and analyte biotin molecules specifically binding to the sensing surface were monitored in real time. The binding of the smallest molecule D-Biotin, with a molecular weight of 244 Da, was easily experimentally observed with a high signal to noise ratio, which shows that the PC-TIR sensor has great potential to be a high-sensitivity and high-throughput sensing technology for small molecule binding analysis.

[1]  X. D. Hoa,et al.  Towards integrated and sensitive surface plasmon resonance biosensors: a review of recent progress. , 2007, Biosensors & bioelectronics.

[2]  Stuart L Schreiber,et al.  Small molecules: the missing link in the central dogma , 2005, Nature chemical biology.

[3]  J. Homola.  Present and future of surface plasmon resonance biosensors , 2003, Analytical and bioanalytical chemistry.

[4]  N. Freeman,et al.  Dual-polarization interferometry: an analytical technique to measure changes in protein structure in real time, to determine the stoichiometry of binding events, and to differentiate between specific and nonspecific interactions. , 2004, Analytical biochemistry.

[5]  Yunbo Guo,et al.  Sensitive molecular binding assay using a photonic crystal structure in total internal reflection. , 2008, Optics express.

[6]  Wolfgang Knoll,et al.  Evaluation of two- and three-dimensional streptavidin binding platforms for surface plasmon resonance spectroscopy studies of DNA hybridization and protein-DNA binding. , 2007, Biosensors & bioelectronics.

[7]  G. Wurtz,et al.  Plasmonic nanorod metamaterials for biosensing. , 2009, Nature materials.

[8]  Shuo Wang,et al.  Sensors and Biosensors for the Determination of Small Molecule Biological Toxins , 2008, Sensors.

[9]  V. Konopsky,et al.  Photonic crystal surface waves for optical biosensing , 2007, 2011 International Workshop on Biophotonics.

[10]  Charles T. Campbell,et al.  Binding and Dissociation Kinetics of Wild-Type and Mutant Streptavidins on Mixed Biotin-Containing Alkylthiolate Monolayers , 2000 .

[11]  Janos Vörös,et al.  Optical microarray biosensing techniques , 2006 .

[12]  Philippe M. Fauchet,et al.  Macroporous Silicon Microcavities for Macromolecule Detection , 2005 .

[13]  W E Moerner,et al.  New directions in single-molecule imaging and analysis , 2007, Proceedings of the National Academy of Sciences.

[14]  Christelle Monat,et al.  Integrated optofluidics: A new river of light , 2007 .

[15]  Yuze Sun,et al.  Sensitive optical biosensors for unlabeled targets: a review. , 2008, Analytica chimica acta.

[16]  L.J. Guo,et al.  Polymer microring resonators for biochemical sensing applications , 2006, IEEE Journal of Selected Topics in Quantum Electronics.

[17]  Herwig Brunner,et al.  Controlled surface functionalization of silica nanospheres by covalent conjugation reactions and preparation of high density streptavidin nanoparticles. , 2004, Journal of nanoscience and nanotechnology.

[18]  Peter B. Dervan,et al.  Regulation of gene expression by small molecules , 1997, Nature.

[19]  J. Homola.  Surface plasmon resonance sensors for detection of chemical and biological species. , 2008, Chemical reviews.

[20]  Jing Yong Ye,et al.  Label-free biosensing using a photonic crystal structure in a total-internal-reflection geometry , 2009, BiOS.

[21]  Sang Jun Sim,et al.  Enhancement of the sensitivity of surface plasmon resonance (SPR) immunosensor for the detection of anti-GAD antibody by changing the pH for streptavidin immobilization , 2004 .

[22]  R. Kunz,et al.  Label-free highly sensitive detection of (small) molecules by wavelength interrogation of integrated optical chips , 2003 .

[23]  Zdenek. Knittl,et al.  Optics of thin films; an optical multilayer theory , 1976 .

[24]  Charles M Lieber,et al.  Label-free detection of small-molecule-protein interactions by using nanowire nanosensors. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[25]  B. Cunningham,et al.  A label-free optical technique for detecting small molecule interactions. , 2002, Biosensors & bioelectronics.