Devices and Methods for Sample Analysis

Abstract

Methods, devices, and systems for analyte analysis using a nanopore are disclosed. The methods, devices, and systems utilize a first and a second binding member that each specifically bind to an analyte in a biological sample. The method further includes detecting and/or counting a cleavable tag attached to the second binding member and correlating the presence and/or the number of tags to presence and/or concentration of the analyte. Certain aspects of the methods do not involve a tag, rather the second binding member may be directly detected/quantitated. The detecting and/or counting may be performed by translocating the tag/second binding member through a nanopore. Devices and systems that are programmed to carry out the disclosed methods are also provided. Also provided herein are instruments that are programmed to operate a cartridge that includes an array of electrodes for actuating a droplet and further includes an electrochemical species sensing region. The instrument may be used to analyse a sample in a cartridge that includes an array of electrodes for actuating a droplet and further includes a nanopore layer for detecting translocation of a tag/second binding member through nanopore. An instrument configured to operate a first cartridge that includes an array of electrodes for actuating a droplet and further includes an electrochemical species sensing region and a second cartridge that includes an array of electrodes for actuating a droplet and further includes a nanopore layer for detecting translocation of a tag/second binding member through nanopore is disclosed. An instrument configured to operate a cartridge that includes an array of electrodes for actuating a droplet, an electrochemical species sensing region, and a nanopore layer for detecting translocation of a tag/second binding member through nanopore is disclosed.

1 - 37 . (canceled) 
     
     
         38 . A system for detection of an analyte in a sample, the system comprising:
 an analyte detection instrument; and   one or more analyte detection cartridges,   wherein the analyte detection instrument comprises:
 a control unit; 
 a detection unit; and 
 a cartridge interface for operable connection to the one or more analyte detection cartridges, 
 wherein the control unit is configured for controlling activation of a plurality of electrodes in a cartridge for moving a droplet present in the cartridge, 
 wherein the detection unit is configured to detect: 
 i) a first analyte related signal from a droplet in a cartridge; and 
 ii) a second analyte related signal from a tag or an analyte-specific binding member translocating through a pore of a nanopore layer in a cartridge, 
 wherein the each one or more analyte detection cartridge comprises: 
 a plurality of electrodes to generate electrical actuation forces on a liquid droplet in response to the control unit; and 
 a detection region for generating an analyte related signal from a droplet in the cartridge; and/or 
 a detection region for generating an analyte related signal from a tag or an analyte-specific binding member translocating through a pore of a nanopore layer in the cartridge. 
   
     
     
         39 . The system of  claim 38 , further comprising reagent reservoirs, wherein the reagent reservoirs are adjacent to the cartridge interface or are located on the cartridge. 
     
     
         40 . The system of  claim 39 , wherein the instrument is programmed for forming droplets from the reagent reservoirs and for moving the droplets by selectively activating and deactivating the plurality of electrodes. 
     
     
         41 . The system of claim  30 , wherein the instrument is programmed for forming at least a sample droplet from a sample introduced into the cartridge and for moving the sample droplet(s) by selectively activating and deactivating the plurality of electrodes. 
     
     
         42 . The system of  claim 38 , wherein the reagent reservoirs comprise a reagent comprising an enzyme specific for the analyte in the sample, wherein the enzyme acts on the analyte to generate a reaction product that generates an electrical or optical signal. 
     
     
         43 . The system of  claim 38 , wherein the reagent further comprises a redox mediator. 
     
     
         44 . The system of  claim 40 , wherein the reagent reservoirs comprise a first binding member that selectively binds to the analyte, wherein the first binding member is attached to a bead. 
     
     
         45 . The system of  claim 44 , wherein the first binding member is an antibody. 
     
     
         46 . The system of  claim 44 , wherein the reagent reservoirs comprise a second binding member that selectively binds to the analyte, wherein the second binding member is attached to an enzyme. 
     
     
         47 . The system of  claim 46 , wherein the second binding member is an antibody. 
     
     
         48 . The system of  claim 46 , wherein the enzyme acts on a substrate to produce a reaction product associated with an electrical signal or an optical signal. 
     
     
         49 . The system of  claim 38 , wherein the cartridge interface comprises an insertion slot for accommodating the cartridge, wherein the cartridge comprises:
 an electrical detection region, the electrical detection region comprising:   a working electrode and a reference electrode for detecting electrical signal from an electrochemical species generated when an analyte is present in the sample; or   a nanopore layer disposed between the first and second substrates, wherein the plurality of electrodes are configured for translocating a tag or an analyte-specific binding member across the nanopore layer and wherein the electrical detection unit detects the translocation.   
     
     
         50 . The system of  claim 49 , wherein the instrument comprises a plurality of insertion slots. 
     
     
         51 . The system of  claim 50 , wherein the plurality of insertion slots are positioned on a carousel. 
     
     
         52 . The system of  claim 38 , wherein the system comprises programming for selecting a function of the instrument, wherein the function is selected based on type of cartridge operably connected to the cartridge interface of the instrument, wherein the function is selected from a plurality of assays for processing the sample. 
     
     
         53 . The system of  claim 52 , wherein when the cartridge comprises an electrical detection region, the instrument is instructed to process the sample for generating an electrical signal in response to presence of an analyte in the sample and to detect the electrical signal via the electrical detection unit. 
     
     
         54 . The system of  claim 53 , wherein instrument receives an indication of the analyte to be detected and processes the sample based on the type of analyte to be detected. 
     
     
         55 . The system of  claim 54 , wherein the indication is provided by a user via a user interface of the instrument or wherein the indication is provided via a machine-readable indicator present on the cartridge. 
     
     
         56 . The system of  claim 38 , wherein when the cartridge comprises an optical detection region, the instrument is instructed to process the sample for generating an optical signal in response to presence of an analyte in the sample and to detect the optical signal via the optical detection unit. 
     
     
         57 . The system of  claim 38 , wherein the system is configured for detection of an analyte in a whole blood sample and/or a plasma fraction of a whole blood sample. 
     
     
         58 . The system of  claim 38 , wherein the system is configured for detection of an analyte in a plasma fraction of a whole blood sample, wherein the plasma fraction of the whole blood sample is generated using a cartridge comprising a filter for separating the plasma fraction from the whole blood sample. 
     
     
         59 - 84 . (canceled)