Self-referencing biodetection method and patterned bioassays
View Patent ↗The invention relates to compositions and methods for detecting biomolecular interactions. The detection can occur without the use of labels and can be done in a high-throughput manner. The invention further relates to self-referencing colorimetric resonant optical biosensors and optical devices.
1. A self-referencing colorimetric resonant optical biosensor comprising:
(a) one or more liquid-holding vessels comprising a colorimetric resonant optical biosensor as a surface; and
(b) one or more specific binding substances immobilized on a first portion of the colorimetric resonant optical biosenor of each liquid-holding vessel forming a reaction surface, and no specific binding substances immobilized on a second portion of the colorimetric resonant optical biosenor of each liquid-holding vessel forming a reference surface, wherein the biosensor comprises two or more reaction surfaces in each liquid holding vessel and two or more reference surfaces in each liquid holding vessel.
2. The self-referencing colorimetric resonant optical biosensor of claim 1 , wherein when the biosensor is illuminated a resonant grating effect is produced on the reflected radiation spectrum and wherein the depth and period of a grating of the biosensor are less than a wavelength of the resonant grating effect.
3. The self-referencing colorimetric resonant optical biosensor of claim 1 , wherein a narrow band of optical wavelengths is reflected from the biosensor when the biosensor is illuminated with a broad band of optical wavelengths.
4. The self-referencing colorimetric resonant optical biosensor of claim 1 , wherein the liquid-holding vessel is selected from the group consisting of a microtiter plate well, a test tube, a Petri dish and a microfluidic channel.
5. A method of detecting the binding of one or more specific binding substances to their respective binding partners in a self-referencing colorimetric resonant optical biosensor of claim 1 comprising:
(a) applying one or more specific binding partners in a reaction volume to the one or more liquid holding vessels of claim 1 ;
(b) illuminating the reaction surfaces and the reference surfaces with light;
(b) detecting a maxima in reflected wavelength, or a minima in transmitted wavelength of light from the reaction surfaces and the reference surfaces; and
(c) comparing the maxima or minima of the one or more reference surfaces to the maxima or minima from the one or more reaction surfaces;
wherein the binding of one or more specific binding substances to their respective binding partners is detected.
6. A method of detecting activity of an enzyme in a self-referencing colorimetric resonant optical biosensor of claim 1 comprising:
(a) applying one or more enzymes in a reaction volume to the one or more liquid holding vessels of claim 1 ;
(b) illuminating the one or more reaction surfaces and the one or more reference surfaces with light;
(c) detecting a maxima in reflected wavelength, or a minima in transmitted wavelength of light from the one or more reaction surfaces and the one or more reference surfaces; and
(d) comparing the maxima or minima of the one or more reference surfaces to the maxima or minima from the one or more reaction surfaces;
wherein the activity of an enzyme is detected.
7. A method of detecting inhibition activity of one or more molecules against one or more enzymes or specific binding partners in a self-referencing colorimetric resonant optical biosensor of claim 1 comprising:
(a) applying one or more molecules suspected of having inhibition activity in a reaction volume to the one or more liquid holding vessels of claim 1 ;
(b) applying one or more enzymes or specific binding partners in a reaction volume to the one or more liquid holding vessels;
(c) illuminating the one or more reaction surfaces and the one or more reference surfaces with light;
(d) detecting a maxima in reflected wavelength, or a minima in transmitted wavelength of light from the one or more reaction surfaces and the one or more reference surfaces; and
(e) comparing the maxima or minima of the one or more reference surfaces to the maxima or minima from the one or more reaction surfaces;
wherein the inhibition activity of one or more molecules is detected.