Semiconductor-based biosensor and detection method therefor

Abstract

To provide a semiconductor-based sensor device and method for detecting a biological substance.SOLUTION: A semiconductor-based sensor 10 has a sensor surface functionalized with detector molecules, and a fluid 16 in contact with the sensor includes an analyte 18 and other species 20. The analyte binds to the detector molecule. Binding of the analyte causes a measurable change in a physical property (e.g., an electrical property) of the semiconductor, and a measured resistance change, ΔR, indicates the presence of the analyte. In this way, the sensor can be used for detecting the concentration of an analyte in a fluid sample.SELECTED DRAWING: Figure 1B

Description

[Technical field]

[0001] Described herein are semiconductor-based sensor devices and methods for detecting biological agents. [Background technology]

[0002] Biosensors generally comprise devices that integrate a binding molecule (also referred to herein as a "detector molecule") and a signal transducer to provide a signal that recognizes the presence of a particular target (e.g., an analyte). Biosensors can be based on a variety of physical principles, but generally rely on the binding of a binding molecule to a target analyte. The specific binding or reaction between the binding molecule and the analyte introduces a signal that can be transmitted and measured. Biosensors can be configured for macromolecular recognition of various types of human cells, viruses, pathogens, and the like. As such, these devices have a wide range of diagnostic utility, including applications in human health, food safety, drug response, and personalized medicine. Summary of the Invention

[0003] Described herein are semiconductor-based sensor devices and methods for detecting biological agents.

[0004] In one aspect, an apparatus is provided comprising a semiconductor sensor functionalized with detector molecules, the apparatus further comprising a heater integrated with the semiconductor sensor and a temperature measurement device integrated with the semiconductor sensor, the heater configured to heat the semiconductor sensor based, at least in part, on a temperature measured by the temperature measurement device.

[0005] In one aspect, a method is provided, the method including measuring a temperature of a portion of an apparatus and / or a fluid in contact with the apparatus, the apparatus comprising a semiconductor sensor functionalized with detector molecules, the method further including heating the semiconductor sensor and / or the fluid, the amount of heat applied to the semiconductor sensor and / or the fluid being based, at least in part, on the temperature measured in the measuring step.

[0006] In some embodiments, the semiconductor sensor comprises a substrate (eg, comprising silicon).

[0007] The heater may comprise a metal wire on the substrate.

[0008] In some embodiments, the temperature measuring device is a micro thermometer.

[0009] In some embodiments, the semiconductor sensor comprises a field effect transistor.The semiconductor sensor may comprise one or more nanowires.

[0010] In some embodiments, the detector molecule is capable of binding an analyte. The detector molecule may comprise, for example, an antibody, a DNA fragment, and / or an RNA fragment. The detector molecule may comprise an enzyme. The detector molecule may comprise glucose oxidase. The analyte may comprise a biomolecule.

[0011] In some embodiments, the step of measuring the temperature includes measuring an electrical characteristic of a temperature measuring device integrated with the semiconductor sensor, the electrical characteristic of the temperature measuring device being selected from the group consisting of resistance, voltage, current, and conductance.

[0012] In some embodiments, the step of heating the semiconductor sensor and / or the fluid includes applying a current or voltage to a heater integrated with the semiconductor sensor, and if the temperature measured during the step of measuring the temperature is equal to or less than a predetermined target temperature, the amount of heat applied to the semiconductor sensor and / or fluid during the step of heating is zero.

[0013] In some embodiments, the method further comprises exposing the semiconductor sensor to a fluid containing an analyte.

[0014] The heating step may be performed such that the temperature measured by the temperature measurement device is within a target temperature range, for example, the target temperature range is from 0° C. to 50° C., and in some cases the target temperature range is from 30° C. to 50° C. The heating step may increase binding of the analyte to the detector molecule compared to binding in the absence of the heating step.

[0015] Other aspects and embodiments will become apparent from the following detailed description of various non-limiting embodiments of the invention, when considered in conjunction with the accompanying figures. In the event that this specification and a document incorporated by reference include conflicting and / or inconsistent disclosure, the specification will control. [Brief description of the drawings]

[0016] Non-limiting embodiments of the present invention are illustratively described with reference to the accompanying figures, which are schematic and are not intended to be drawn to scale, in which each identical or nearly identical component shown is typically represented by a single numeral, and for clarity, not every component is labeled in every figure, nor are every component of each embodiment of the present invention shown unless illustration is necessary for a person skilled in the art to understand the invention. [Figure 1A] FIG. 1A shows a schematic diagram of an exemplary semiconductor sensor in the absence of analyte binding to the detector molecule, according to an embodiment. [Figure 1B] FIG. 1B shows a schematic diagram of an exemplary semiconductor sensor in the presence of analyte binding to a detector molecule, according to an embodiment. [Diagram 2] FIG. 2 shows a schematic diagram of a semiconductor sensor with an integrated heater and an integrated temperature measurement device according to an embodiment. [Diagram 3] FIG. 3 shows a schematic diagram of a semiconductor sensor with an integrated heater and an integrated temperature sensor, according to one non-limiting embodiment. [Figure 4] FIG. 4 shows a non-limiting design of a device with a semiconductor sensor having an integrated heater and an integrated temperature sensor, according to one non-limiting embodiment. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0017] Described herein are semiconductor-based sensor devices and methods for detecting biological materials (e.g., analytes). The devices may be used to detect analytes in a fluid in contact with the device. The analytes may bind to detector molecules functionalized on the surface of the sensor. Such binding may result in a measurable signal to detect the presence of the analyte. As described further below, certain embodiments include controlling the temperature of the fluid and / or the sensor device to affect the binding of the analyte to the detector molecules. For example, the sensor may be integrated with a heater and a temperature measurement device (e.g., a temperature sensor such as a thermometer) to control the temperature of the binding site above room temperature to increase the binding strength and, therefore, improve detectability. Advantageously controlling the temperature also allows for better control of the Debye length. (For example, increasing the temperature may increase the Debye length, increasing the signal measured by the sensor and improving detectability.) The sensors may be used to detect the presence of specific molecules that may be useful in food safety, drug reactions, personalized medicine, cancer detection, disease validation, and other medical and biological applications.

[0018] 1A-1B show a semiconductor-based sensor 10 according to one embodiment. Detector molecules 12 are functionalized on a surface 14 of the sensor. As described below, the detector molecules are selected for their ability to bind to an analyte one wishes to detect. FIG. 1B shows a fluid 16 in contact with the sensor. As shown, the fluid contains an analyte 18 and other species 20. In this embodiment, the analyte binds to the detector molecule. Binding of the analyte causes a measurable change in a physical property (e.g., an electrical property) of the semiconductor. For example, in an exemplary embodiment, as shown in FIGS. 1A and 1B, a measured change in resistance ΔR indicates the presence of the analyte. In this manner, the sensor can be used to detect the concentration of an analyte in a fluid sample.

[0019] It should be understood that changes in physical properties other than resistance may be measured. For example, in some embodiments, changes in conductance (or ΔG) may be measured, and in some embodiments, changes in conductivity may be measured. In some embodiments, a conformational change upon binding of the detector molecule causes a measurable change. In some embodiments, the change is due to electrical gating by the analyte. In some embodiments, the change is due to a change in surface plasmon resonance. In some embodiments, the change in the property (e.g., conductance, resistance) may be detected electrically, typically by applying a current to the sensor and measuring the change in voltage. In some embodiments, the change in the property (e.g., conductance, resistance) may be detected electrically, typically by applying an alternating electric field.

[0020] Any suitable semiconductor-based sensor device may be used. For example, suitable devices are described in commonly owned U.S. patent application Ser. No. 14 / 510,178, filed Oct. 9, 2014, and commonly owned International Application No. PCT / US2015 / 041527, published as International Publication No. WO2016 / 089453, all of which are incorporated herein by reference in their entirety.

[0021] The sensor comprises a semiconductor material. Suitable semiconductor materials from which nanosensors can be fabricated include, but are not limited to, silicon, germanium, III-V semiconductors, and the like. In some embodiments, the sensor comprises silicon. The sensor may include one or more layers formed above or below the semiconductor material, such as additional layers of semiconductor material or insulating layers (e.g., oxides such as silicon oxide).

[0022] In some embodiments, the semiconductor is patterned into nanowire(s). The sensor may be a field effect transistor sensor (e.g., a nanosensor) in some embodiments. In general, a field effect transistor (FET) uses an electric field to control a conducting channel and, therefore, the conductivity of charge carriers in the channel. By applying an electric field to the gate to change the size and shape of the conducting channel, the flow of charge carriers between the source and drain can be tuned. In an exemplary biosensor configuration, the FET comprises a nanosensor (e.g., nanowire) channel between the source and drain terminals. The surface of the nanosensor (e.g., nanowire) can be biofunctionalized such that biomolecular binding events can create an electric field similar to the controlled electric field applied to a conventional FET (FIG. 1). In some devices using FET principles, a specified and physically separated sensor surface can be formed by precision fabrication. The FET sensor can be connected to electronic circuitry to monitor the specific conductance of the sensor surface. In some embodiments, in operation, many independent electronic circuits may be interrogated in a massively parallel manner. FET biosensors can be adapted for the measurement of biomolecules that interact with such sensor surfaces (FIGS. 1A and 1B).

[0023] In some embodiments, the sensor comprises a nanoscale silicon-based FET device. Many such devices exhibit the sensitivity, reliability, robustness, and sensor flexibility required for many multiplexed diagnostic microarrays. In some cases, nanoscale devices can be developed and / or implemented on traditional top-down silicon. In some cases, developing and implementing nanoscale devices on traditional top-down silicon can improve the reliability and robustness of top-down silicon semiconductor manufacturing processes and reduce test error rates, both at the point of care and in central laboratories. This, in some cases, can increase the effectiveness of each patient visit to a laboratory or clinic, reduce diagnostic costs, and enable earlier diagnosis, treatment, and monitoring.

[0024] In certain embodiments, the nanosensor is a silicon nanochannel field effect transistor (FET) biosensor. Such sensors can be used for sensitive and / or label-free detection of analytes. Such sensors can have excellent electrical properties and small dimensions. In certain embodiments, silicon nanochannels are ideally suited for ultra-high sensitivity. In some cases, the high surface-to-volume ratio of these systems allows for single molecule detection. In some cases, such FET sensors (e.g., biosensors) offer the advantages of high-speed, low-cost, high-yield fabrication without sacrificing the sensitivity typical of traditional optical approaches in diagnostics. Using top-down fabrication methods, the advantages of complementary metal oxide semiconductor (CMOS) technology can be exploited to realize richly multiplexed sensor arrays. Examples of nanochannel-based sensor systems are described, for example, in International Patent Publication WO 2008 / 063901 A1 by Yucheng et al., and International Patent Publication WO 2009 / 124111 A1 by Mohanty et al., each of which is incorporated by reference in its entirety for all purposes.

[0025] In some embodiments, the semiconductor-based sensor can be part of a bias and measurement circuit. In some embodiments, the bias and measurement circuit operates by applying a bias voltage across the nanosensor (e.g., nanochannel) in the circuit. The bias voltage can be selected to be sufficiently negative to achieve a desired dependence of the differential conductance of the sensing element on the surface potential of the sensor (e.g., surface potential of the nanochannel). In some embodiments, this dependence has a steep slope region of high gain that is substantially greater than the reference gain exhibited by the sensing element in a zero bias state, thereby achieving a relatively high signal-to-noise ratio. The bias and measurement circuit, in some embodiments, measures the differential conductance of the sensing element and converts the measured differential conductance into a signal indicative of the presence or activity of the analyte. In some embodiments, the measured differential conductance can be converted into a signal indicative of the presence or activity of the analyte by using a look-up table or alternative conversion mechanism that reflects a previous calibration operation. In some embodiments, the applied gate voltage can be used to control the sensitivity of the sensor. According to some embodiments, the bias gate voltage and the reference gate voltage can be used independently to control the sensitivity.

[0026] As described above, at least a portion (e.g., a surface) of the sensor is functionalized with a detector molecule. The detector molecule may include, for example, an antibody, an enzyme, a protein, a peptide, a small molecule, a nucleic acid, an aptamer, a receptor molecule, a polymer, and / or a supramolecular structure. The detector molecule may be designed to be particle specific, i.e., only one particular analyte binds to a particular detector molecule. In some embodiments, the detector molecule is an antibody. In some embodiments, the detector molecule is or includes a DNA or RNA fragment. In some embodiments, the detector molecule is or includes glucose oxidase. In some cases, the detector molecule is or includes urease.

[0027] Analytes may include, for example, proteins, small molecules, nucleic acids, peptides, antibodies, aptamers, biomarkers, genes, virus particles, supramolecular structures, macromolecules, receptor molecules, biological cells, and / or biological cell clusters. In some cases, the analyte includes a biomolecule. In some examples, the analyte may be a protein biomarker or a genetic biomarker. In some embodiments, the analyte is or includes a monosaccharide and / or polysaccharide. For example, in some cases, the analyte is or includes glucose. One non-limiting example of a small molecule analyte is urea.

[0028] It is to be understood that the methods described herein are not limited to a particular detector molecule or analyte.

[0029] In some embodiments, the analyte and detector molecules may be bonded through a chemical interaction, such as a chemical bond. The chemical bond may be a covalent bond or a non-covalent bond. In some cases, the chemical bond is a non-covalent bond, such as a hydrogen bond, an ionic bond, a coordinate bond, and / or a van der Waals interaction. One or more of the species and / or substances (e.g., analyte, detector molecule) may include a functional group capable of forming such a bond. It should be understood that covalent and non-covalent bonds between components may be formed by any type of reaction, with appropriate functional groups to undergo such a reaction, as known to those of skill in the art. Chemical interactions suitable for use in the various embodiments described herein can be readily selected by those of skill in the art based on the description herein.

[0030] In some embodiments, the association between the analyte and detector species may occur through a biological binding phenomenon (i.e., between a pair of complementary biological molecules). For example, the analyte or detector molecule may contain an entity such as biotin that specifically binds to a complementary entity such as avidin or streptavidin on another species or substance. Other examples of biological molecules that may form biological bonds between pairs of biological molecules include, but are not limited to, proteins, nucleic acids, glycoproteins, carbohydrates, hormones, etc. Non-limiting examples include, but are not limited to, antibody / peptide pairs, antibody / antigen pairs, enzyme / substrate pairs, enzyme / inhibitor pairs, enzyme / cofactor pairs, protein / substrate pairs, nucleic acid / nucleic acid pairs, protein / nucleic acid pairs, peptide / peptide pairs, protein / protein pairs, small molecule / protein pairs, receptor / hormone pairs, receptor / effector pairs, ligand / cell receptor pairs, biotin / avidin pairs, biotin / streptavidin pairs, drug / target pairs, small molecule / peptide pairs, small molecule / protein pairs, and small molecule / enzyme pairs, etc. Biological interactions between species and / or substances(s) for use in the embodiments described herein can be readily selected by one of skill in the art based on the description herein of their functions, examples of such biological interactions, and knowledge in the art of straightforward techniques for identifying suitable biological interactions.

[0031] In certain embodiments, the analyte and detector species may be associated with one another through physical interactions, for example, in some embodiments, the analyte (e.g., a supramolecular structure) may be physically entangled with at least a portion of the detector species (e.g., a macromolecule).

[0032] In certain embodiments, the analyte and detector species may be associated with one another via a linking moiety (e.g., another biological or chemical species that brings the analyte and detector species into close proximity). For example, the minimum distance between the analyte and detector species that are associated with one another may be greater than a Debye length. In some examples, the minimum distance may be about 100 nanometers or less, about 50 nanometers or less, about 25 nanometers or less, about 10 nanometers or less, or about 1 nanometer or less.

[0033] As discussed above, certain embodiments include controlling the temperature of the fluid and / or sensor device to affect binding of the analyte to the detector molecule. The temperature can be controlled by a heating step that enhances binding of the analyte to the detector molecule compared to the absence of the heating step. For example, without being bound to a particular theory, in some cases, when the fluid and / or sensor are at a relatively low temperature, the analyte may bind relatively poorly to the detector molecule (e.g., due to a temperature dependence of the binding constant or due to a temperature dependent structure of the analyte and / or detector molecule). In such cases, the heating step can increase the temperature of the fluid and / or sensor, thereby enhancing binding of the analyte to the detector molecule (e.g., by increasing the temperature dependent binding constant or by causing a conformational change in the analyte and / or detector molecule that results in a structure more conducive to binding).

[0034] 2 shows a schematic diagram of a semiconductor-based sensor 10 having an integrated heater 22 and an integrated temperature measurement device 24, according to one embodiment. The sensor also includes detector molecules 12 functionalized on a sensor surface 14 as described above. As shown diagrammatically, the heater and temperature measurement device are formed on the sensor surface in close proximity to the detector molecules such that the temperature of the binding sites can be controlled to be at a selected value and / or within a selected range, according to one embodiment.

[0035] FIG. 3 illustrates another embodiment in which the sensing region 26 of the sensor is within a region 28 that contains a fluid sample 30. Region 28 may be, for example, a microfluidic chamber. In this embodiment, the heater 22 is located near the end of region 28 and the temperature measuring device is located on the opposite side of the heater. It should be understood that other arrangements of the heater and temperature measuring device may be used to provide appropriate temperature control. For example, one or both of the heater and / or temperature measuring device may be located within region 28. In such an embodiment, it may be important to take care to avoid interference between the heater current and the voltage applied to the sensor and / or fluid. Some embodiments utilize multiple heaters and / or temperature measuring devices.

[0036] The heater 22 may be any suitable heating element that may be integrated with a sensor. In some embodiments, the heater comprises an elongated structure such as a wire (e.g., a metal wire). For example, a serpentine metal wire may be deposited on a substrate of different conductivity (e.g., an insulating substrate). A current may be passed through the wire to generate Joule heating that causes an increase in temperature of the substrate. The temperature may be controlled by controlling the current passing through the wire.

[0037] In some embodiments, the heater comprises a thermoelectric material that converts voltage into heat. By controlling the voltage across the thermoelectric material, the temperature may be controlled.

[0038] It should also be appreciated that other suitable heaters may be used.

[0039] The temperature measuring device 24 may be any suitable such device that can be integrated with a sensor. In some cases, the temperature measuring device is a micro-thermometer. In some embodiments, the temperature measuring device is a material whose resistivity is temperature dependent, such as a thin film metal or semiconductor. In such embodiments, the temperature is measured by measuring the resistance. In some embodiments, the temperature measuring device is a thermoelectric material that generates a measured voltage in response to a change in temperature.

[0040] It should also be appreciated that other suitable heaters may be used.

[0041] Generally, any suitable method for controlling the heater using a temperature measurement device can be used. In some embodiments, an output signal from a temperature measurement device can be used to control an input signal to the heater. For example, the heater can be activated by passing a current through the heater (e.g., a metal wire) and the temperature can be measured using a thermometer (e.g., an integrated micro thermometer). The current or voltage output driving the heater can be adjusted using a feedback loop until the temperature stabilizes at the target temperature and / or range. In some cases, the temperature is controlled to a target temperature range of 0°C to 50°C. In some cases, the target temperature range can be 30°C to 50°C. In some cases, the temperature is controlled to an accuracy of within + / - 5°C, and in some cases within + / - 2°C.

[0042] The following are non-limiting examples illustrating certain features of the sensors described herein. EXAMPLES

[0043] This example illustrates the formation of a sensor that includes an integrated heater and temperature measurement device, as shown in FIG.

[0044] The sensor is made by overlaying a thin silicon layer, typically about 100 nm thick, on a silicon dioxide layer, typically about 200 nm thick.

[0045] The nanowires and electrode attachment pads are then created by electron beam lithography and reactive ion etching, which removes the silicon everywhere except in the sensor area. In some embodiments, this is split into two steps, and the nanowires and pads are defined separately. In some embodiments, the nanowires are defined by electron beam lithography, and the pads are defined by photolithography. In some embodiments, the pads and nanowires are all defined by photolithography.

[0046] A pattern of metal electrodes in the four-point measurement geometry is then photolithographically created and the metal electrodes are deposited. The electrodes can be metals such as Au, Cu, Ag, Al, or alloys or metal multilayers. In some cases, adhesion layers of Ti, Ta, or other metals are used. The electrodes are about 10-20 microns wide near the sensor and become wider the further away. Typically the metal thickness is in the 100 nm range.

[0047] The electrodes are then coated with a thick insulating barrier, typically about 100 nm, typically an oxide such as Al2O3, SiO2, or HfO2.

[0048] The nanowires and electrodes are then coated with a thin top insulating barrier (typically 10 nm) made of a similar oxide.

[0049] Finally, metal pads, as shown in FIG. 4, are deposited to allow connection to external measurement and control equipment. The pads are typically 1-2 microns thick and made of highly conductive noble metals such as Au, Ag, Cu, etc. In some embodiments, utilizing a metallic heater / thermometer system, the heater and thermometer are deposited simultaneously with the pads, with the connections shown in the figure. In other embodiments, the heater and thermometer are placed at a later step.

[0050] A schematic of the completed sensor, ready for integration into the final circuit, is shown in Figure 4. This picture includes additional pads that in some embodiments connect to a reference electrode and other control mechanisms. The liquid to be analyzed is placed in an oval shaped area centered on the sensor.

Claims

1. a semiconductor-based sensor functionalized with a detector molecule; a heater integrated with the semiconductor-based sensor; 1. An apparatus comprising: a temperature measurement device integrated with a semiconductor-based sensor, the heater being configured to heat the semiconductor-based sensor based, at least in part, on a temperature measured by the temperature measurement device.

2. The apparatus of claim 1 , wherein the semiconductor-based sensor comprises a substrate.

3. The apparatus of claim 1 , wherein the heater comprises a metal wire on a substrate.

4. The apparatus of claim 1 , wherein the temperature measuring device is a micro thermometer.

5. The apparatus of claim 1 , wherein the semiconductor-based sensor comprises a field effect transistor.

6. The apparatus of claim 1 , wherein the semiconductor-based sensor comprises one or more nanowires.

7. The device of claim 1 , wherein the detector molecule is capable of binding an analyte.

8. The apparatus of claim 1 , wherein the detector molecule comprises an antibody, a DNA fragment, and / or an RNA fragment.

9. The apparatus of claim 1 , wherein the detector molecule comprises an enzyme.

10. The device of claim 1 , wherein the detector molecule comprises glucose oxidase.

11. The device of claim 1 , wherein the detector molecule comprises urease.

12. The apparatus of claim 7 , wherein the analyte comprises a biomolecule.

13. The apparatus of claim 7 , wherein the analyte comprises a carbohydrate.

14. The apparatus of claim 7 , wherein the analyte comprises glucose.

15. The device of claim 7 , wherein the analyte comprises urea.

16. measuring the temperature of a part of the device and / or a fluid in contact with said device, comprising a semiconductor-based sensor functionalized with detector molecules; and heating the semiconductor-based sensor and / or the fluid, wherein an amount of heat applied to the semiconductor-based sensor and / or the fluid is based, at least in part, on the temperature measured in the measuring step.

17. The method of claim 16 , wherein the step of measuring the temperature includes measuring an electrical characteristic of a temperature measurement device integrated with the semiconductor-based sensor.

18. 20. The method of claim 17, wherein the electrical characteristic of the temperature measuring device is selected from the group consisting of resistance, voltage, current, and conductance.

19. The method of claim 16 , wherein the step of heating the semiconductor-based sensor and / or the fluid comprises applying a current or voltage to a heater integrated with the semiconductor-based sensor.

20. 17. The method of claim 16, wherein the amount of heat applied to the semiconductor-based sensor and / or fluid during the heating step is zero if the temperature measured during the temperature measuring step is equal to or less than a predetermined target temperature.

21. 20. The method of claim 16, comprising exposing the semiconductor-based sensor to a fluid containing an analyte.

22. The method of claim 16 , wherein the semiconductor-based sensor comprises a substrate.

23. 20. The method of claim 19, wherein the semiconductor-based sensor comprises a substrate and the heater comprises a metal wire on the substrate.

24. The method of claim 17 , wherein the temperature measuring device is a micro thermometer.

25. The method of claim 16 , wherein the semiconductor-based sensor comprises a field effect transistor.

26. The method of claim 16 , wherein the semiconductor-based sensor comprises one or more nanowires.

27. The method of claim 21 , wherein the detector molecule is capable of binding the analyte.

28. The method of claim 16 , wherein the detector molecules include antibodies, DNA fragments, and / or RNA fragments.

29. The method of claim 16 , wherein the detector molecule comprises an enzyme.

30. The method of claim 16 , wherein the detector molecule comprises glucose oxidase.

31. The method of claim 16 , wherein the detector molecule comprises urease.

32. The method of claim 21 , wherein the analyte comprises a biomolecule.

33. 22. The method of claim 21 , wherein the analyte comprises a carbohydrate.

34. 22. The method of claim 21 , wherein the analyte comprises glucose.

35. The method of claim 21 , wherein the detector molecule comprises urease.

36. 17. The method of claim 16, wherein the heating step is performed such that the temperature measured by a temperature measuring device is within a target temperature range.

37. 37. The method of claim 36, wherein the target temperature range is from 0°C to 50°C.

38. 37. The method of claim 36, wherein the target temperature range is from 30°C to 50°C.

39. 22. The method of claim 21, wherein the heating step increases binding of the analyte to the detector molecule compared to binding in the absence of the heating step.

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