Multiplexed Assay Using Spectrally-Encoded Solid Support Matrices

Abstract

In a multiplexed assay, each molecule of a plurality of molecules is attached to a support matrix particle with a substrate adapted for attachment and/or synthesis of molecules. A spectrally-encoded identifier embodied in a photochemical medium is embedded or encased within the substrate to uniquely identify the molecule attached to the substrate. The molecules are exposed to one or more processing conditions, and then placed within the path of an optical detector adapted to read the spectrally-encoded identifier and measure biochemical activity on each support matrix particle. The measured biochemical activity is associated with the unique identity of the support matrix particle and, hence, with the molecule attached to the particle.

1 . A multiplexed assay, comprising:
 attaching each molecule of a plurality of molecules to a separate support matrix particle, each support matrix particle comprising a substrate adapted for attachment of a molecule and a spectrally-encoded identifier embodied in a photochemically-active medium incorporated into the substrate, the spectrally-encoded identifier adapted to uniquely identify the molecule attached to the substrate;   exposing the plurality of molecules and their corresponding support matrix particles to one or more processing conditions in suspension to produce a plurality of processed molecules;   placing the plurality of support matrix particles into a vessel feeding into an optically-transparent tube disposed within the detection path of each of an optical detector and an analytical instrument, wherein the optical detector comprises a light source emitting at a wavelength adapted to induce activity in the photochemically-active medium for reading the spectrally-encoded identifier and wherein the analytical instrument is adapted to measure biochemical activity on each support matrix particle; and   associating the measured biochemical activity with the spectrally-encoded identifier for the corresponding support matrix particle and, hence, with the processed molecule that is associated with the spectrally-encoded identifier.   
     
     
         2 . The assay of  claim 1 , wherein the photochemically-active medium is a photochromic material. 
     
     
         3 . The assay of  claim 1 , wherein the optically-transparent tube causes the support matrix particles to flow past the optical detector one at a time. 
     
     
         4 . The assay of  claim 1 , wherein the light source comprises a laser. 
     
     
         5 . The assay of  claim 1 , wherein the spectrally-encoded identifier comprises a plurality of different wavelengths. 
     
     
         6 . The assay of  claim 1 , wherein the one or more processing conditions comprises attaching a fluorescent label to each molecule. 
     
     
         7 . The assay of  claim 6 , wherein the analytical instrument measures biochemical activity by detecting an emission from the fluorescent label. 
     
     
         8 . The assay of  claim 1 , wherein the one or more processing conditions comprise:
 reacting the molecules with an analyte; and   further reacting the molecules with a detection antibody, wherein the detection antibody is conjugated with a detectable label.   
     
     
         9 . The assay of  claim 8 , wherein the analyte comprises a fluid comprising an antigen. 
     
     
         10 . The assay of  claim 1 , wherein the spectrally-encoded identifier is embedded or encased within the substrate. 
     
     
         11 . The assay of  claim 1 , wherein the spectrally-encoded identifier is encased within a protective coating. 
     
     
         12 . The assay of  claim 1 , wherein a remote memory stores the encoded data in association with information about the molecule attached to the support matrix particle. 
     
     
         13 . An assay for evaluating a plurality of molecules, comprising:
 linking each of the molecules to a separate support matrix particle having a spectrally-encoded identifier embodied in a photochemical medium within the support matrix particle, the spectrally-encoded identifier having a unique identity, wherein the unique identity is associated with the molecule linked to the support matrix particle;   exposing each molecule and its linked support matrix particle to one or more reagents in suspension, wherein at least one of the one or more reagents is conjugated with a label;   placing the plurality of support matrix particles into a vessel feeding into an optically-transparent tube disposed within the path of an optical detector comprising a light source for inducing activity in the photochemical medium and a photodetector adapted to read the spectrally-encoded identifier as the support matrix particle passes through the optically-transparent tube; and   measuring biochemical processes on each support matrix particle by directing the support matrix particles past an analytical instrument adapted to detect the label.   
     
     
         14 . The assay of  claim 13 , wherein the support matrix particle has a fluor incorporated therein. 
     
     
         15 . The assay of  claim 14 , wherein the spectrally-encoded identifier is embedded or encased within the support matrix. 
     
     
         16 . The assay of  claim 13 , wherein the spectrally-encoded identifier comprises a plurality of different wavelengths. 
     
     
         17 . The assay of  claim 13 , wherein the label is a fluorescent label. 
     
     
         18 . The assay of  claim 7 , wherein the step of measuring comprises detecting an emission from the fluorescent label. 
     
     
         19 . The assay of  claim 13 , wherein the photochemical medium is a photochromic material. 
     
     
         20 . The assay of  claim 13 , wherein the optically-transparent tube causes the support matrix particles to flow past the optical detector one at a time. 
     
     
         21 . The assay of  claim 13 , wherein the light source comprises laser. 
     
     
         22 . An assay system, comprising:
 a plurality of support matrix particles, each support matrix particle adapted for linking a molecule of a plurality of molecules, wherein each support matrix particle has a spectrally-encoded identifier embodied in a photochemical medium embedded or encased therein, the spectrally-encoded identifier having a unique identity, wherein the unique identity is associated with the molecule linked to the support matrix particle;   a particle identity reader adapted for reading the spectrally-encoded identifier for each support matrix particle and generating a signal corresponding to the spectrally-encoded identifier;   an analytical instrument adapted to measure biochemical activity on each support matrix particle and generate a signal corresponding to measured biochemical activity;   a vessel adapted for receiving a plurality of support matrix particles with linked molecules and directing the plurality of support matrix particles within a reading field of the particle identity reader and a detection field of the analytical instrument; and   a system controller in communication with the particle identity reader and the analytical instrument for receiving the signal corresponding to the spectrally-encoded identifier and the signal corresponding to measured biochemical activity, wherein the system controller is operable to associate the measured biochemical activity with the spectrally-encoded identifier for the corresponding support matrix particle and, hence, with the molecule that is linked to the spectrally-encoded identifier.   
     
     
         23 . The assay system of  claim 22 , wherein the plurality of molecules are labeled with a fluorescent label and the analytical instrument comprises an excitation light source and a fluorescence detector. 
     
     
         24 . The assay system of  claim 22 , wherein the particle identity reader is adapted to detect a plurality of different wavelengths. 
     
     
         25 . The assay system of  claim 22 , further comprising an optically-transparent tube in fluid communication with the vessel, wherein the optically-transparent tube directs the plurality of support matrix particles past the particle identity reader and the analytical instrument. 
     
     
         26 . The assay system of  claim 22 , wherein the vessel comprises a plate 
     
     
         27 . An assay for evaluating a plurality of molecules, comprising:
 linking each of the molecules to a separate support matrix particle having a spectrally-encoded identifier comprising a photochemically-active medium embedded or encased therein, the spectrally-encoded identifier having a unique identity that is associated with the molecule linked to the support matrix particle;   exposing the plurality of molecules and their corresponding support matrix particles to one or more reagents to produce processed molecules;   placing a plurality of support matrix particles with linked processed molecules into a common vessel;   measuring biochemical processes within the common vessel using analytical instrumentation;   before, during or after the step of measuring, detecting the spectrally-encoded identifier for each support matrix particle, wherein the spectrally-encoded identifier comprises a plurality of different wavelengths; and   associating the measured biochemical process for each support matrix particle with the spectrally-encoded identifier for that particle and, hence, with the processed molecule that is associated with the spectrally-encoded identifier.   
     
     
         28 . The assay of  claim 27 , wherein the support matrix particle further comprises a magnetic material. 
     
     
         29 . The assay of  claim 27 , wherein the common vessel comprises a plate. 
     
     
         30 . The assay of  claim 27 , wherein the step of exposing comprises attaching a detectable label, and the step of measuring comprises:
 exciting the detectable label with a light source; and   detecting fluorescent emissions from the detectable label.