Portable instrument for in vitro detection and quantification of biomarkers
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Publication Number
US-10908156-B2
Publication Date
2021-02-02
Expiration Date
Abstract
A portable assay instrument for rapid detection and quantification of biomarkers is described. The instrument may include components to automate steps of an assay. Assay-specific agitation during an incubation phase in which capture binding agents, detection binding agents, and conjugates are present in sample wells can appreciably increase the speed of the assay and improve assay sensitivity. The instrument may further include an optical detector and processor for automated probing of the sample wells and computation of biomarker concentration.
Core Innovation
The invention provides a method for detecting and quantifying a target biomarker with an integrated instrument. A single-use and disposable prepared sample well is sealed prior to being received in a sample holder of the integrated instrument, and the sealed well contains an immobilized binding agent specific to the target biomarker, a detection binding agent specific to the target biomarker in a solution, a blocking solution comprising at least one of bovine serum albumin (BSA) and non-fat dry milk, and a stability buffer capable of preserving and enhancing assay components over time.
After receiving a sample in the prepared sample well, the integrated instrument automatically agitates the sample well at an agitation station, detects radiation from the sample well with a detector of the integrated instrument, and quantifies a concentration of the target biomarker based on the detected radiation. The method includes timing constraints in which the agitation period is less than 10 minutes and a time from initiation of agitation and calculation of the target biomarker concentration is no greater than 30 minutes.
The document further refines radiation-based quantification by defining illuminations and detector-based calculations for determining biomarker concentration from optical readouts. In particular embodiments, illumination with light at two different characteristic wavelengths and absorbance comparisons between the characteristic wavelengths are used for quantifying the concentration of the target biomarker, including an approach that can rely on a subtractive comparison.
Claims Coverage
The independent claim recites an integrated-instrument workflow with a sealed disposable prepared sample well, automatic agitation, radiation detection, and timing limits. Dependent claims refine the readout and add specific structural or chemical details for the sample well, including a membrane seal, specific capture binding agent identity, casein-based blocking, two-wavelength absorbance comparison, and limiting the target biomarker to chronic transplant rejection or cancer.
Integrated instrument biomarker detection and quantification
Automatically agitating the sample well at an agitation station of the integrated instrument; detecting radiation from the sample well with a detector of the integrated instrument; and quantifying a concentration of the target biomarker based on the detected radiation from the sample well.
Sealed single-use disposable prepared sample well with preserved assay components
Receiving a single-use and disposable prepared sample well in a sample holder of the integrated instrument, wherein prior to being received in the sample holder the single-use and disposable prepared sample well is sealed to contain an immobilized binding agent specific to the target biomarker; a detection binding agent specific to the target biomarker in a solution; a blocking solution comprising at least one of bovine serum albumin (BSA) and non-fat dry milk; and a stability buffer capable of preserving and enhancing assay components over time.
Rapid agitation and concentration calculation timing constraints
Wherein the agitation period is less than 10 minutes and wherein a time from initiation of agitation and calculation of the target biomarker concentration is no greater than 30 minutes.
Two-wavelength absorbance comparison quantification
Illuminating the sample well with light at two different characteristic wavelengths and quantifying the concentration of a target biomarker by comparing absorbance values measured at those wavelengths.
Chronic transplant rejection or cancer biomarker indication
Using a target biomarker that indicates chronic transplant rejection or cancer.
Overall, the claim set centers on a sealed disposable prepared sample well integrated into an instrument that performs automatic agitation, radiation detection, and rapid concentration calculation, with further refinements including two-wavelength absorbance comparison quantification and restricting the target biomarker to chronic transplant rejection or cancer.
Stated Advantages
Improved assay sensitivity/time response as described by prototype performance using agitation conditions.
Rapid assay completion, with total time from agitation initiation to concentration calculation no greater than 30 minutes as recited.
Achieves sensitivity of about 66 pg/mL as reported in prototype results.
Documented Applications
Detecting and quantifying VEGF-C in contexts including chronic transplant rejection and cancer, as described in the document’s target biomarker contexts.
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