A label-free optical technique for detecting small molecule interactions.
A novel approach for the label-free detection of molecular interactions is presented in which a colorimetric resonant grating is used as a surface binding platform. The grating, when illuminated with white light, is designed to reflect only a single wavelength. When molecules are attached to the surface, the reflected wavelength (color) is shifted due to the change of the optical path of light that is coupled into the grating. By linking receptor molecules to the grating surface, complementary binding molecules can be detected without the use of any kind of fluorescent probe or radioactive label. The detection technique is capable of detecting the addition and removal of small molecules as they interact with receptor molecules on the sensor surface or enzymes in the solution surrounding the sensor. Two assays are presented to exemplify the detection of small molecule interactions with the biosensor. First, an avidin receptor layer is used to detect 244 Da biotin binding. Second, a protease assay is performed in which a 136 Da p-nitroanilide (pNA) moeity is cleaved from an immobilized substrate. Because the sensor structure can be embedded in the plastic surfaces of microtiter plates or the glass surfaces of microarray slides, it is expected that this technology will be most useful in applications where large numbers of biomolecular interactions are measured in parallel, particularly when molecular labels will alter or inhibit the functionality of the molecules under study. Screening of pharmaceutical compound libraries with protein targets, and microarray screening of protein-protein interactions for proteomics are examples of applications that require the sensitivity and throughput afforded by this approach.
Lin
Bo
B
SRU Biosystems, 14A Gill Street, Woburn, MA 01801, USA.
Qiu
Jean
J
Gerstenmeier
John
J
Li
Peter
P
Pien
Homer
H
Pepper
Jane
J
Cunningham
Brian
B
D001360:Avidin / Q000378:metabolism*; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D001710:Biotin / Q000378:metabolism*; D053148:Caspase 3; D020169:Caspases / Q000032:analysis / Q000378:metabolism*; D003124:Colorimetry / Q000295:instrumentation / Q000379:methods; D004789:Enzyme Activation; D004800:Enzymes, Immobilized / Q000378:metabolism; D046911:Macromolecular Substances; D055095:Optics and Photonics / Q000295:instrumentation*; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000295:instrumentation / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013057:Spectrum Analysis / Q000295:instrumentation / Q000379:methods*; D013379:Substrate Specificity; D017186:Titrimetry / Q000295:instrumentation / Q000379:methods
D004800:Enzymes, Immobilized; D046911:Macromolecular Substances; D001360:Avidin; D001710:Biotin; D053148:Caspase 3; D020169:Caspases