DISPERSION INJECTION METHODS FOR BIOSENSING
First Claim
1. An injection method for determining interaction parameters between an analyte and a ligand in a biosensor, the biosensor comprising a flow channel conduit in fluid communication with a flow cell conduit, said flow cell conduit housing at least a first and second sensing region, said first sensing region having a ligand immobilized thereon, the method comprising the steps of:
- obtaining a fluid sample containing an analyte having a starting concentration;
injecting the fluid sample through the flow channel conduit under physical injection conditions sufficient to cause the analyte to undergo a defined dispersion event in route to the flow cell conduit thereby creating an analyte concentration gradient, wherein the analyte concentration gradient comprises a maximum analyte concentration and a minimum analyte concentration that differ by at least one order of magnitude;
measuring the responses elicited by the analyte interacting with the ligand at the first sensing region as the analyte concentration gradient progresses continuously through the flow cell conduit, wherein the measured responses provide a response curve;
incorporating a dispersion term into an interaction model, wherein the dispersion term represents a gradient profile generated by the defined dispersion event thereby providing the analyte concentration present at the first sensing region at any time point during the injection;
anddetermining the interaction parameters by fitting the interaction model to the response curve.
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Abstract
I Injection methods for determining biomolecular interaction parameters such in label-free biosensing systems are provided. The methods generally relate to analyte sample injection methods that generate well-defined analyte concentration gradients en route to a sensing region possessing an immobilized binding partner. The injections conditions are generally established according to a set of rules that create a dispersion event that can be accurately modeled by a dispersion term. The dispersion term is incorporated into the desired interaction model to provide a reliable representation of the analyte concentration gradient profile, The resulting interaction model is then fitted to a measured binding response curve in order to calculate the interaction parameters. Thus, the injection methods described herein provide a continuous analyte titration allowing a full analyte dose response to be recorded in a single injection in contrast to the standard multiple injection approach
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Citations
41 Claims
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1. An injection method for determining interaction parameters between an analyte and a ligand in a biosensor, the biosensor comprising a flow channel conduit in fluid communication with a flow cell conduit, said flow cell conduit housing at least a first and second sensing region, said first sensing region having a ligand immobilized thereon, the method comprising the steps of:
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obtaining a fluid sample containing an analyte having a starting concentration; injecting the fluid sample through the flow channel conduit under physical injection conditions sufficient to cause the analyte to undergo a defined dispersion event in route to the flow cell conduit thereby creating an analyte concentration gradient, wherein the analyte concentration gradient comprises a maximum analyte concentration and a minimum analyte concentration that differ by at least one order of magnitude; measuring the responses elicited by the analyte interacting with the ligand at the first sensing region as the analyte concentration gradient progresses continuously through the flow cell conduit, wherein the measured responses provide a response curve; incorporating a dispersion term into an interaction model, wherein the dispersion term represents a gradient profile generated by the defined dispersion event thereby providing the analyte concentration present at the first sensing region at any time point during the injection; and determining the interaction parameters by fitting the interaction model to the response curve. - View Dependent Claims (6, 7, 9, 10, 11, 16, 18, 20, 21, 32, 33, 34, 35, 36)
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37. An injection method for determining interaction parameters between an analyte and a ligand in a biosensor, the biosensor comprising a flow channel conduit in fluid communication with a flow cell conduit, said flow cell conduit housing at least a first and a second sensing region, said first sensing region having a ligand immobilized thereon, the method comprising the steps of:
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obtaining a fluid sample containing an analyte having a starting concentration; injecting the fluid sample through the flow channel conduit under physical injection conditions sufficient to cause the analyte to undergo dispersion in route to the flow cell conduit thereby creating an analyte concentration gradient, wherein the analyte concentration gradient comprises a maximum analyte concentration and a minimum analyte concentration that differ by at least one order of magnitude, and wherein the physical injection conditions are consistent with those necessary to produce Taylor dispersion; measuring the responses elicited by the analyte interacting with the ligand at the first sensing region as the analyte concentration gradient progresses continuously through the flow cell conduit, wherein the measured responses provide a response curve; incorporating a dispersion term into an interaction model, wherein the dispersion term represents a gradient profile consistent with Taylor dispersion; and determining the interaction parameters by fitting the interaction model to the response curve. - View Dependent Claims (38)
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39. A computer program for calculating kinetic or binding parameters of an interaction between an analyte and a ligand in a biosensing system comprising:
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program code for an interaction model including a dispersion term, wherein the dispersion term is incorporated into the interaction model as the concentration of the analyte injected, wherein the dispersion term encodes a gradient profile produced by dispersion conditions during the injection; program code for fitting a response curve produced by the interaction between the analyte and ligand to the interaction model; and program code for calculating the kinetic or binding parameters of the interaction. - View Dependent Claims (40, 41)
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Specification