Free solution measurement of molecular interactions by backscattering interferometry
First Claim
1. A method for free-solution determination of molecular interactions comprising the steps of:
- (a) providing a substrate having a channel formed therein for reception of a fluid sample to be analyzed;
(b) introducing a first sample comprising a first non-immobilized analyte to be analyzed into the channel;
(c) introducing a second sample comprising a second non-immobilized analyte to be analyzed into the channel;
(d) allowing the first analyte to interact with the analyte to form one or more interaction products;
(e) directing a coherent light beam onto the substrate such that the light beam is incident on the channel to generate backscattered light through reflective and refractive interaction of the light beam with a substrate/channel interface and the sample, the backscattered light comprising interference fringe patterns including a plurality of spaced light bands whose positions shift in response to changes in the refractive index of the fluid sample;
(f) detecting positional shifts in the light bands; and
(g) determining an equilibrium constant of the interaction from the detected shifts.
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Abstract
Disclosed are methods, systems, and apparatuses for the free solution measurement of molecular interactions by backscattering interferometry (BSI). Molecular interaction can be detected between analytes in free-solution wherein at least one of the analytes is label-free and detection is performed by back-scattering interferometry. Further, molecular interaction can be detected between analytes in free-solution, wherein at least one of the analytes is label-free, wherein one of the analytes is present in a concentration of less than about 5.0×10−7M. Also disclosed are label-free, free-solution, and/or real-time measurements of characteristic properties and/or chemical events using the disclosed techniques. The disclosed methods can have very low detection limits and/or very low sample volume requirements. Also disclosed are various biosensor applications of the disclosed techniques. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present invention.
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Citations
22 Claims
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1. A method for free-solution determination of molecular interactions comprising the steps of:
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(a) providing a substrate having a channel formed therein for reception of a fluid sample to be analyzed; (b) introducing a first sample comprising a first non-immobilized analyte to be analyzed into the channel; (c) introducing a second sample comprising a second non-immobilized analyte to be analyzed into the channel; (d) allowing the first analyte to interact with the analyte to form one or more interaction products; (e) directing a coherent light beam onto the substrate such that the light beam is incident on the channel to generate backscattered light through reflective and refractive interaction of the light beam with a substrate/channel interface and the sample, the backscattered light comprising interference fringe patterns including a plurality of spaced light bands whose positions shift in response to changes in the refractive index of the fluid sample; (f) detecting positional shifts in the light bands; and (g) determining an equilibrium constant of the interaction from the detected shifts. - View Dependent Claims (2, 3, 4, 5, 6, 7, 9, 10, 11)
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8. The method of clam 1, wherein the substrate and channel together comprise a capillary tube.
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12. A method for free-solution determination of molecular interactions comprising the steps of:
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(a) providing a substrate having a channel formed therein for reception of a fluid sample to be analyzed; (b) introducing a first sample comprising a first non-immobilized analyte to be analyzed into the channel; (c) introducing a second sample comprising a second non-immobilized analyte to be analyzed into the channel; (d) allowing the first analyte to interact with the second analyte to form one or more interaction products; (e) directing a coherent light beam onto the substrate such that the light beam is incident on the channel to generate backscattered light through reflective and refractive interaction of the light beam with a substrate/channel interface and the sample, the backscattered light comprising interference fringe patterns including a plurality of spaced light bands whose positions shift in response to changes in the refractive index of the fluid sample; (f) detecting positional shifts in the light bands; and (g) determining an end-point value of the interaction from the detected shifts. - View Dependent Claims (13, 14, 15, 16, 17, 18, 19, 20, 21, 22)
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Specification