Method and system for biomarker detection

Abstract

Method, system, and kit of parts for electrochemical analysis of aldehydes in biological samples. A trapping agent comprising an aniline and / or an aniline derivative is added to the biological sample and electrochemical testing is performed on the sample. In some embodiments, the trapping agent is 4-aminobiphenyl. In some embodiments, the testing is performed using a portable testing system comprising a container with premixed solution including the trapping agent, an electrochemical sensor, a plurality of electrodes, and electronics that control a voltage / current / charge applied to the solution for testing. The trapping agent may be bound to the surface of one or more electrodes. The concentration of the aldehyde biomarker(s) can then be determined using a connected data processing device and shared with a user of the portable testing system. The testing system may detect multiple biomarkers. The testing system may detect multiple biomarkers simultaneously.

Description

METHOD AND SYSTEM FOR BIOMARKER DETECTIONCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to US Provisional App. No. 63 / 612,786, entitled “CONCUSSION DIAGNOSIS SYSTEM AND METHOD”, filed December 20, 2023, and to US Provisional App. No. 63 / 665,501, entitled “METHOD AND SYSTEM FOR BIOMARKER DETECTION”, filed June 28, 2024, the entire disclosures of which are incorporated herein by reference.TECHNICAL FIELD

[0002] The present invention relates to biomarker detection. More specifically, the present invention relates to detection of aldehydes in biological samples.BACKGROUND

[0003] Biomarkers are specific compounds within biological samples that may indicate various conditions. Further, increases and decreases of such biomarkers may indicate changes in these conditions that are of interest. For example, various diseases and conditions are associated with increases in oxidative stress. Oxidative stress induces lipid peroxidation, and thereby produces various aldehyde species, aldehyde compounds, etc. Accordingly, aldehydes are one possible group of interesting biomarkers.

[0004] For example, recent studies using animal models indicate that aldehyde concentrations increase after concussive brain trauma, including traumatic brain injury (TBI) and specifically mild TBI (mTBI). mTBI is known to be difficult to detect, relying largely on self-reported symptoms, responses to questionnaires, and other qualitative assessments such as reflex responses. Existing quantitative approaches are expensive, time-consuming, and do not generally convey information about mTBI. For example, the protein-based blood test offered by Abbott Laboratories requires a CT-scan and is generally only used to rule out severe brain injury. Aldehydes are relatively small molecules, with several reports in the literature confirming that aldehydes can cross the blood-brainbarrier. In comparison, other biomarkers proposed for detection of concussion (such as the glial fibrillary acidic protein (GFAP) used by Abbott Laboratories) are large molecules which might have limited abilities to cross this barrier, hindering applications of these sensors. A quantitative approach based on the increase of an aldehyde biomarker would thus be of significant interest to healthcare providers, patients with mTBI, and others.

[0005] Aldehydes may also be relevant biomarkers for a variety of other conditions, including, without limitation, Alzheimer’s disease, Parkinson’s disease, cardiovascular disease, nonalcoholic fatty liver disease (NAFLD), cancer, diabetes, chronic obstructive pulmonary disease (COPD), kidney disease, Huntington’s disease (HD), amyotrophic lateral sclerosis (ALS), and bronchitis. There is thus a significant interest in being able to obtain clear quantitative measurements of aldehyde concentrations in biological samples.

[0006] However, to date, quantitative measurement of aldehyde concentrations has been difficult. In particular, aldehydes are not significantly electrochemically active on their own. There is therefore a need for low-cost and rapid methods and systems for quantitative (essentially electrochemical) analysis of aldehydes in biological samples.SUMMARY

[0007] This document discloses a method, system, and kit of parts for electrochemical analysis of aldehydes in biological samples. A trapping agent comprising an aniline and / or an aniline derivative is combined with the biological sample and electrochemical testing is performed on the sample. In some embodiments, the trapping agent is 4-aminobiphenyl. In some embodiments, the testing is performed using a portable testing system comprising a container with premixed solution including the trapping agent, an electrochemical sensor, a plurality of electrodes, and electronics that control a voltage / current / electrical charge applied to the solution for testing. The trapping agent may be bound to the surface of one or more electrodes. The concentration of the aldehyde biomarker(s) can then be determined using a connected data processing deviceand shared with a user of the portable testing system. The testing system may detect multiple biomarkers.

[0008] In a first aspect, this document discloses a method for detecting levels of a biomarker in a biological sample, said method comprising: receiving said biological sample; combining a trapping agent with said biological sample, said trapping agent being selected to enhance an electrochemical response of said at least one biomarker in said biological sample; evaluating said electrochemical response; and based on said electrochemical response, determining a concentration of said at least one biomarker in said biological sample, wherein said biomarker is an aldehyde, and wherein said trapping agent comprises at least one of an aniline and an aniline derivative.

[0009] In another embodiment, this document discloses a method wherein said trapping agent comprises at least one of 4-aminobiphenyl (4AB) and anthranilic acid (AAcid).

[0010] In another embodiment, this document discloses a method wherein said concentration correlates to a severity of a condition of a human patient.

[0011] In another embodiment, this document discloses a method wherein said condition is at least one of: a concussion, traumatic brain injury (TBI), and / or mild traumatic brain injury (mTBI); Alzheimer’s disease; Parkinson’s disease; cardiovascular disease; nonalcoholic fatty liver disease (NAFLD); cancer; diabetes; chronic obstructive pulmonary disease (COPD); kidney disease; Huntington’s disease (HD); amyotrophic lateral sclerosis (ALS); and bronchitis.

[0012] In another embodiment, this document discloses a method further comprising the step of detecting a concentration of at least one other biomarker in said biological sample.

[0013] In another embodiment, this document discloses a method wherein said at least one other biomarker and said aldehyde are simultaneously detected.

[0014] In another embodiment, this document discloses a method wherein said evaluating step is performed using a portable electrochemistry device.

[0015] In another embodiment, this document discloses a method further comprising displaying an indication relating to said concentration to a user of said device.

[0016] In another embodiment, this document discloses a method wherein said evaluating step uses at least one of square wave voltammetry, chronoamperometry, and chronopotentiometry.

[0017] In another embodiment, this document discloses a method wherein said biological sample is at least one of: blood; saliva; urine; sweat; and cerebrospinal fluid (CSF).

[0018] In a second aspect, this document discloses a system for detecting levels of a biomarker in a biological sample, said system comprising: a container for said biological sample and for a solution, said solution comprising a trapping agent selected to enhance an electrochemical response of said at least one biomarker in said biological sample; a contact-based sensor for sensing said electrochemical response when said biological sample is mixed with said solution in said container, said sensor being within said container and in contact with said mixture; a plurality of electrodes coupled to said sensor for applying one of a voltage and a current to said sensor and for receiving said electrochemical response through said sensor; and an electronics module for controlling said one of said voltage and said current for measuring characteristics of said electrochemical response, wherein said biomarker is an aldehyde, wherein said trapping agent comprises at least one of an aniline and an aniline derivative, wherein said electronics module interfaces with a data processing device to thereby allow said data processing device to analyze said electrochemical response, and wherein said electronics module is coupled to said plurality of electrodes to allow for control of said one of said voltage and said current and for reception and measurement of said electrochemical response.

[0019] In another embodiment, this document discloses a system wherein said trapping agent comprises at least one of 4-aminobiphenyl (4AB) and anthranilic acid (AAcid).

[0020] In another embodiment, this document discloses a system wherein said concentration correlates to a severity of a condition of a human patient.

[0021] In another embodiment, this document discloses a system wherein said condition is at least one of: a concussion, traumatic brain injury (TBI), and / or mild traumatic brain injury (mTBI); Alzheimer’s disease; Parkinson’s disease; cardiovascular disease; nonalcoholic fatty liver disease (NAFLD); cancer; diabetes; chronic obstructive pulmonary disease (COPD); kidney disease; Huntington’s disease (HD); amyotrophic lateral sclerosis (ALS); and bronchitis.

[0022] In another embodiment, this document discloses a system wherein the system is configured to detect a concentration of at least one other biomarker in said biological sample.

[0023] In another embodiment, this document discloses a system wherein said at least one other biomarker and said aldehyde are simultaneously detected.

[0024] In another embodiment, this document discloses a system wherein said biological sample is at least one of: blood; saliva; urine; sweat; and cerebrospinal fluid (CSF).

[0025] In another embodiment, this document discloses a system wherein said trapping agent is bound to a surface of at least one electrode of said plurality of electrodes.

[0026] In a third aspect, this document discloses a kit of parts for detecting levels of a biomarker in a biological sample, said kit of parts comprising: a container for said biological sample and for a solution, said solution comprising a trapping agent selected to enhance an electrochemical response of said at least one biomarker in said biological sample; a contact-based sensor for sensing said electrochemical response when said biological sample is mixed with said solution in said container, said sensor being for use within said container and in contact with said mixture; a plurality of electrodes coupled to said sensor for applying one of a voltage and a current to said sensor and for receiving said electrochemical response through said sensor; and an electronics module for controlling said one of said voltage and said current and for measuring characteristics of said electrochemical response, wherein said biomarker is an aldehyde, wherein said trapping agent comprises at least one of an aniline and an aniline derivative, wherein said electronics module interfaces with a data processing device to thereby allow said data processing device to analyze saidelectrochemical response, and wherein said electronics module is coupled to said plurality of electrodes to allow for control of said one of said voltage and said current and for reception and measurement of said electrochemical response.

[0027] In another embodiment, this document discloses a kit of parts wherein said trapping agent comprises at least one of 4-aminobiphenyl (4AB) and anthranilic acid (AAcid).

[0028] In another embodiment, this document discloses a kit of parts wherein said trapping agent is bound to a surface of at least one electrode of said plurality of electrodes.

[0029] In another embodiment, this document discloses a kit of parts further comprising a collection apparatus for obtaining said biological sample.BRIEF DESCRIPTION OF THE DRAWINGS

[0030] The present invention will now be described by reference to the following figures, in which identical reference numerals refer to identical elements and in which:Figure 1 is a flowchart detailing a method according to an aspect of the invention;Figure 2 is a structure diagram of exemplary trapping agents, specifically AAcid and 4 AB;Figures 3A and 3B show electrochemical responses of different concentrations of a trapping agent (AAcid) in stock solution;Figures 3C and 3D show electrochemical responses of different concentrations of a trapping agent (4AB) in stock solution;Figure 4 shows the effect(s) of various conditions on voltammograms during experimental optimization procedures;Figure 5 shows voltammograms taken at multiple time intervals with the trapping agent 4 AB;Figure 6A shows voltammograms of the electrochemical response of 4AB with different concentrations of butyraldehyde;Figure 6B shows voltammograms of the electrochemical response of 4AB with different concentrations of MDA;Figures 7A and 7B show peak voltages and standard curves based on the butyraldehyde response in Figure 6A;Figures 8A to 8C show peak voltages and standard curves based on the MDA response in Figure 6B;Figure 9A shows voltammograms of multiple biogenic aldehyde species with 4 AB;Figure 9B shows voltammograms of multiple ketone species with 4AB; andFigure 10 shows an exemplary process in which the method and system disclosed herein may be used.DETAILED DESCRIPTION

[0031] To better understand the present invention, the reader is directed to the listing of citations at the end of this description. For ease of reference, these citations and references have been referred to by their listing number throughout this document. The contents of the citations in the list at the end of this description are hereby incorporated by reference herein in their entirety.

[0032] The present invention provides methods and systems for identifying biogenic aldehydes (i.e., aldehydes naturally occurring in biological samples). In particular, the present invention provides a rapid, accurate, portable, and quantitative sensor for detecting aldehydes and determining the concentrations thereof in biological samples. As noted above, aldehydes may provideindications of and / or useful information relating to numerous conditions, including without limitation Alzheimer’s disease, Parkinson’s disease, cardiovascular disease, nonalcoholic fatty liver disease (NAFLD), cancer, diabetes, chronic obstructive pulmonary disease (COPD), kidney disease, Huntington’s disease (HD), amyotrophic lateral sclerosis (ALS), and bronchitis.

[0033] Biogenic aldehydes include, without limitation, butyraldehyde, malondialdehyde (MDA), 4-hydroxyneonal (4HNE), decadienal, formaldehyde, acetaldehyde, acrolein, glyoxal, methylglyoxal, crotonaldehyde, hexanal, heptanal, 4- hydroxyhexenal (4-HHE), 3-aminopropanal, and 3,4- dihydroxyphenylacetaldehyde (DOPAL). Given that these aldehydes are not significantly electrochemically active, as noted above, detecting their presence and / or concentration in biological samples can be difficult. Accordingly, a trapping agent is combined with the sample. As should be understood, the trapping agent and sample may be combined in another (preferably relatively inert or neutral) solution. The trapping agent is selected to be electrochemically active and to bond with the aldehyde(s), thereby enhancing the electrochemical response of the aldehyde and facilitating detection.

[0034] Once the trapping agent has been combined with the sample, the electrochemical response of the sample is evaluated. As will be described in more detail below, this electrochemical evaluation / testing is, in some embodiments, performed using a portable testing system or kit. In other embodiments, the biological samples may be sent for testing at a lab and / or tested using conventional testing equipment. Based on the test results, that is, the electrochemical response of the biological sample, the concentration of the biomarker(s) in the biological sample is determined.

[0035] This process is detailed in Figure 1. At step 100, the sample is received. At step 110, the trapping agent is added to / combined with the sample. At step 120, the electrochemical response of the sample is evaluated. At step 130, based on the electrochemical response, the concentration of the aldehyde biomarker(s) is determined.

[0036] The aldehyde biomarker may be any of the aldehydes listed above and / or other relevant aldehydes depending on the implementation. Multiple biomarkers may be simultaneously detected. The multiple biomarkers may include multiple aldehydes. As well, the multiple biomarkers may include one or more aldehyde biomarkers and one or more non-aldehyde biomarkers, such as GFAP and / or ubiquitin C-terminal hydrolase LI (UCH-L1). Such multiplex testing provides simultaneous determination of multiple different biomarkers, each of which may convey different information regarding the biological samples.

[0037] The trapping agents comprise anilines and / or aniline derivatives. For example, the trapping agent may be a conjugated amine such as 4-aminobiphenyl (4AB). Other aniline derivatives may also be used, including without limitation anthranilic acid (AA or AAcid). The structures of AAcid and 4AB are shown in Figure 2.Experimentation and Testing

[0038] Various experimental tests were conducted to evaluate the proposed method. A stock solution of dimethyl sulfoxide (DMSO), a non-toxic, non-volatile solvent with a wide electrochemical window was used in testing. As DMSO is non- conductive, an electrolyte (specifically tetrabutylammonium perchlorate, TBAP) was added. Different stock solutions in different implementations, of course, may yield slightly different results — it is known, for example, that the relative intensities of various peaks in voltammograms varies slightly between different days and different stock solutions.

[0039] Figure 3A is a chart showing experimental tests of different concentrations of trapping agent AAcid in stock solution. The left chart shows the concentration response in the oxidative sweep and the right chart shows the concentration response in the reduction sweep. For AAcid, the optimal performance was determined to occur at 0.5mM (based on the closest response to the example scan of 0.5mM AAcid + 0.5mM butyraldehyde). Figure 3B, similarly, shows the electrochemical response of AAcid with different concentrations of representative analytes (aldehyde) and a possible competing reagent (ketone).

[0040] Figure 3C is a chart showing experimental tests of different concentrations of trapping agent 4AB in stock solution. The left chart shows the concentration response in the oxidative sweep and the right chart shows the concentration response in the reduction sweep. For 4AB, the optimal performance was determined to occur at 0.25mM (based on the closest response to the example scan of 0.25mM 4AB + 0.5mM butyraldehyde). Figure 3D, similarly, shows the electrochemical response of 4AB with representative analytes (aldehyde) and a possible competing reagent (ketone).

[0041] Note that the trapping agent-analyte reactions are 1:1. As such, and noting that aldehydes are not typically present in large concentrations, smaller concentrations of trapping agent are generally preferred, as they permit analysis of biologically relevant levels of analyte. However, the concentration of the combined trapping agent and aldehyde must also be sufficient to reproducibly generate detectable voltammograms. As such, it should be understood that the “optimal” concentrations disclosed above may be better suited to some implementations than to others.

[0042] As well, it was noted that 4AB produced more consistent and reproducible baseline voltammograms than AAcid (compare Figure 3C to Figure 3A), and had a clean, reproducible response with increasing concentration of the trapping agent. Further, upon the addition of butyraldehyde, a dramatic response was observed in both the oxidative and reductive sweeps for 4AB, providing multiple potential peaks for analysis. Additionally, as seen when comparing of Figure 3B to Figure 3D, AAcid showed a similar magnitude of response for both aldehydes and ketones (in other tests), which indicates that AAcid is not highly selective for aldehydes. 4AB, in contrast, showed a larger and more variable response to aldehydes than to ketones (see Figure 3D). As such, many other tests were conducted using 4 AB.

[0043] Depending on the embodiment, the electrochemical testing may use any suitable voltammetry method that may be known in the art. In some embodiments, the electrochemical response of the biological samples is tested using square wave voltammetry (SWV). A powerful feature of SWV is that multiple parameters can be altered to optimize the electrochemical response of a specific analyte,including (but not limited to) frequency, amplitude, pulse magnitude, and window size. These parameters can be adjusted individually and during the testing process (i.e., with live monitoring and adjustment to maximize the response while minimizing the signal to noise ratio). Figure 4 shows the effect of various conditions on the SWV voltammograms, in experimental optimizations. (Again, the oxidative sweep is at left and the reduction sweep is at right.)

[0044] It should be noted that other electrochemical procedures may be used to assess the electrochemical response(s) of the sample. As non-limiting examples, the testing may use chronoamperometry, chronopotentiometry, and / or other pulsed voltammetry procedures, as well as any suitable combination of procedures.

[0045] During testing, reaction times of the reagents were also evaluated. As should be understood, if the electrochemical response is evaluated too early after mixing the biological samples with the testing solution (i.e., stock solution + trapping agent), the reagents will not have had time to come to equilibrium and the results will not accurately reflect the concentration(s) of biomarker(s). Based on sampling a testing solution of stock solution of equimolar 4AB (0.25mM) with butyraldehyde, a reaction time of approximately 20 minutes was found to provide a reproducible maximum in current response. Figure 5 shows voltammograms taken at multiple time intervals during this test process. Of course, the optimal time may be different in different implementations, depending on the concentrations used and reagents sought. However, 20 minutes is a significant improvement over existing, time-consuming detection methods currently available.

[0046] To determine the concentration range at which the biosensor platform used in testing could detect aldehydes, samples of various aldehyde concentrations for both the model aldehyde (butyraldehyde) and a biogenic aldehyde (MDA) were prepared. Butyraldehyde samples ranging from 0.0625mM up to 3mM were prepared and analyzed with 0.25mM 4AB, using the testing conditions described above. As is shown in Figure 6A, a clear relationship between maximum current achieved and aldehyde concentration was observed. Both an oxidation peak (centered at -0. IV) and a reduction peak (at approximately -0.2V) were observedto be affected by increasing concentration of butyraldehyde. Additionally, the shape of the oxidation peaks shifted slightly above a certain concentration of butyraldehyde. The presence of multiple unique features in both sweeps affords the possibility of using multiple analysis points, allowing for greater sensitivity and specificity in the final biosensor.

[0047] An identical experiment was performed using MDA, a biogenic aldehyde, with a slightly different concentration range of 0.125mM to 4mM (Figure 6B). As with butyraldehyde, a dramatic current response was observed in both the oxidation and reduction sweeps, with the behavior even more pronounced with this biogenic aldehyde than with butyraldehyde. While the major oxidative and reductive peaks were found in the same locations as butyraldehyde, they did have slightly different peak shapes. Interestingly, a third region in the MDA voltammogram was identified as providing concentration information. Between 0.4V and 0.9V, a change from two individual peaks to a single peak was observed when going from low to high MDA concentrations.

[0048] Peak voltages and currents could be calculated based on the Figure 6A and Figure 6B data. (Note that the peak currents are possibly correlated to concentration of analytes.) The response of butyraldehyde, shown in Figures 7A and 7B, was much simpler than that of MDA (shown in Figures 8A to 8C and discussed more below). The butyraldehyde response displayed a single dominant peak (compared to the baseline), appearing at -0.02V. For this aldehyde, simply taking the magnitude of this peak provided a suitable standard curve (shown at right in Figure 7A). A secondary analysis technique, integrating the area under the curve of the first peak, was also performed (left of Figure 7B); this produced a standard curve (right of Figure 7B) with similar features to the non-integrated curve of Figure 7 A.

[0049] As noted above, the voltammograms for MDA were more complex than those for butyraldehyde. The MDA voltammograms (shown in Figure 6B) showed three distinct peaks that appeared and degraded depending on concentration, requiring a more sophisticated analysis method to extend the range of concentrations. The simple “current at one potential” approached used for butyraldehyde, using the magnitude of the peak at -0.2V, yielded poor resolution in the lowerconcentration ranges (see Figure 8A). Further, integrating the area under the curve to capture all the peaks (Figure 8B) did not yield a satisfactory result. However, an algorithm was developed that added or subtracted the magnitude of the different peaks depending on whether the first peak crossed a certain threshold (specifically, if Peakl < 6mV, then Peakl - Peak2 + Peak3; and if Peakl< 6mV, then Peakl + Peak2 + Peak3). This provided a linear standard curve which had excellent slope and linear fit values resulting in strong quantification at low concentrations (Figure 8C). (Dashed lines in Figures 8A to 8C are the 95% confidence interval.)

[0050] Figure 9A shows voltammograms of multiple different biogenic aldehyde species (i.e., 4HNE and decadi enal in addition to the earlier tested MDA and butyraldehyde) to determine whether they could be distinguished from one another and / or from ketone counterparts with similar structures (specifically, from acetone, butanone, and decanone, shown in Figure 9B). As can be seen in Figure 9A, each aldehyde resulted in positive current increase, and each had a unique voltammogram. Note that none of the ketones investigated during this study had as significant of an electrochemical response compared to these aldehydes, and in particular to MDA, especially in the reduction sweep. This indicates a high degree of selectivity towards aldehydes.Analysis System and Device

[0051] As mentioned above, the testing may be performed using a portable sensor device, system, or kit, similar to the system described in US Patent Application No. 18 / 289,469, the entirety of which is hereby incorporated herein by reference. That is, in one aspect, the present invention comprises a system comprising a container / sample tube filled with a testing solution (for example, a 15 mL centrifuge tube). The container also receives the biological sample when collected. The system further comprises a sensor (such as a sensor cap for the container / tube), the sensor comprising a plurality of electrodes for transmitting a voltage / current / charge / etc. into the solution and measuring a current response from the solution and an electronics module / reader device that interfaces between the sensor / sensor cap and a data processing device that analyzes data received from the electronics module. Of course, as would be understood, thechemical makeup of the testing solution depends on the desired implementation (e.g., the biomarker(s) to be identified, etc.). Further, the electrodes may be any suitable electrodes comprising any suitable materials. For example, the electrodes may comprise gold electrodes. Alternatively and / or additionally, the electrodes may comprise carbon electrodes, such as graphene electrodes. All other suitable electrode types and materials should be understood to be within the scope of the invention.

[0052] As well, the trapping agent(s) may be added to the testing solution, present in the sample tube before the testing solution is added, and / or bound to the surface of one or more of the electrodes. Any suitable means of combining the trapping agent with the biological sample fall within the scope of the invention.

[0053] Further, the system may also include collection apparatus for collecting the biological samples. This collection apparatus may include, without limitation, one or more lancets, hypodermic needles and syringes, collection containers, and / or swabs, depending on the nature of the biological sample intended to be assessed. Depending on the embodiment, the biological samples may include blood or blood plasma, urine, saliva, sweat / perspiration, cerebrospinal fluid (CSF), and / or other bodily fluids.

[0054] In some embodiments, the electronics module displays results to a user by way of the data processing device (which may be a user’s personal data device, such as (without limitation) a tablet, smartphone, laptop or desktop computer, etc.). Alternatively and / or additionally, the electronics module displays results to a user through a display, screen, or indicator on the module itself. In some embodiments, the data processing device and the electronics module are contained within a single physical unit.

[0055] Further, the ‘results’ can be understood as indications relating to the determined biomarker concentration(s). The indications may be numerical data (i.e., actual concentration data). Such data may be preferred by sophisticated users of the testing system, such as healthcare professionals. Indications of the concentrations may also be displayed categorically — for example, by displaying the words “low”, “medium”, or “high” — or may be correlated to conditionseverity or likely prognosis. Other categorizations may of course be used as necessary. Additionally, indicators such as coloured lights (e.g, coloured LEDs) may also be used to indicate a general category of results. Such a display format may be preferred by less-sophisticated users of the testing system, such as individuals, school teachers, sports coaches, etc. Indications may also include indicators corresponding to negative or inconclusive results. Other types of indications may also be imagined and fall within the scope of the invention.

[0056] An advantage of point-of-care biomarker analysis performed using a portable testing system is that the analysis can be performed by those with no advanced scientific training and / or outside of clinical settings. Further, as the analysis methods described herein are relatively quick to perform (as noted above, some concentrations of 4AB have been determined to have optimal response times as low as 20 minutes), the system and methods also providing useful results and quick information to healthcare professionals in clinical settings, in contrast with current qualitative analyses and more complex and time-consuming assessments, especially in the context of traumatic brain injury.

[0057] The elements of the system can also be supplied as a kit of parts for assembly by the user. In such kits, the testing solution is preferably supplied in a prefdled tube with the selected trapping agent already added (e.g, with the trapping agent already in the testing solution and / or bound to one or more of the electrodes). Further, each individual test is preferably already self-calibrated. In such cases, the user need only add the biological sample, position the sensor, and connect the tube / container to the electronics module. However, in some embodiments, the various elements and chemical compounds may be supplied in bulk and mixed by the user at the time of testing. Such kits, as should be clear, may not be suitable for use by those without scientific or medical training.Exemplary Process of Use

[0058] Figure 10 depicts atypical assessment process, in which aldehyde concentration is evaluated post-head-injury. In this process diagram, following a head trauma and (potential) concussion, the injured individual visits a clinician. A blood sample is collected and combined with a prepared solution in a test kit of aportable testing device. Results of the test / indications relating to the biomarker concentration are then displayed to the user(s) (i.e., the clinician). Again, it should be understood that this process diagram is merely exemplary, and that aldehyde concentration may convey useful information for a variety of conditions.References

[0059] As noted above, for a better understanding of the present invention, the following references may be consulted. Each of these references is hereby incorporated by reference in its entirety:[1] Marchand, D. K.; Severn, M., Biomarker-Based Point-of-Care Tests for the Evaluation of Mild Traumatic Brain Injury. Ottawa: CADTH; 2020 Mar (CADTH Issues in Emerging Health Technologies, 188).[2] Canadian Patent Application No. 3,052,505; Shuhendler, A. et al.; UNIVERSITY OF OTTAWA; Methods and compounds for detection and binding of aldehydes.[3] International Patent Application PCT / US2018 / 027353; McQuiston, B. et al; ABBOTT LABORATORIES; Methods for aiding in the hyperacute diagnosis and determination of traumatic brain injury in a human subject using early biomarkers.

[0060] As used herein, the expression “at least one of [x] and [y]” means and should be construed as meaning “[x], [y], or both [x] and [y]”

[0061] It should be clear that various components of the present invention, in particular analysis components implemented on the data processing device and / or the electronics module, may be implemented as software modules in an overall software system. As such, components of the present invention may thus take the form of computer executable instructions that, when executed, implement various software modules with predefined functions.

[0062] Such components of the invention may be executed by a computer processor or similar device programmed in the manner of method steps, or may be executedby an electronic system which is provided with means for executing these steps. Similarly, an electronic memory means such as computer diskettes, CD-ROMs, Random Access Memory (RAM), Read Only Memory (ROM) or similar computer software storage media known in the art, may be programmed to execute such method steps. As well, electronic signals representing these method steps may also be transmitted via a communication network.

[0063] Components of the invention may be implemented in any conventional computer programming language. For example, some components may be implemented in a procedural programming language (e.g, “C” or “Go”) or an object-oriented language (e.g, “C++”, “java”, “PHP”, “PYTHON” or “C#”). Further, some components of the invention may be implemented as pre-programmed hardware elements, other related physical components, or as a combination of hardware and software components.

[0064] Components of the invention can be implemented as a computer program product for use with a computer system. Such implementations may include a series of computer instructions fixed either on a tangible medium, such as a computer readable medium (e.g, a diskette, CD-ROM, ROM, or fixed disk) or transmittable to a computer system, via a modem or other interface device, such as a communications adapter connected to a network over a medium. The medium may be either a tangible medium (e.g, optical or electrical communications lines) or a medium implemented with wireless techniques (e.g, microwave, infrared or other transmission techniques). The series of computer instructions embodies all or part of the functionality previously described herein. Those skilled in the art should appreciate that such computer instructions can be written in a number of programming languages for use with many computer architectures or operating systems. Furthermore, such instructions may be stored in any memory device, such as semiconductor, magnetic, optical or other memory devices, and may be transmitted using any communications technology, such as optical, infrared, microwave, or other transmission technologies. It is expected that such a computer program product may be distributed as a removable medium with accompanying printed or electronic documentation (e.g, shrink-wrapped software), preloaded with a computer system (e.g, on systemROM or fixed disk), or distributed from a server over a network (e.g., the Internet or World Wide Web). Of course, some embodiments of the invention may be implemented as a combination of both software (e.g., a computer program product) and hardware. Still other embodiments of the invention may be implemented as entirely hardware, or entirely software (e.g, a computer program product).

[0065] A person understanding this invention may now conceive of alternative structures and embodiments or variations of the above all of which are intended to fall within the scope of the invention as defined in the claims that follow.

Claims

We claim:

1. A method for detecting levels of a biomarker in a biological sample, said method comprising:(a) receiving said biological sample;(b) combining a trapping agent with said biological sample, said trapping agent being selected to enhance an electrochemical response of said at least one biomarker in said biological sample;(c) evaluating said electrochemical response; and(d) based on said electrochemical response, determining a concentration of said at least one biomarker in said biological sample, wherein said biomarker is an aldehyde, and wherein said trapping agent comprises at least one of an aniline and an aniline derivative.

2. The method according to claim 1, wherein said trapping agent comprises at least one of 4-aminobiphenyl (4AB) and anthranilic acid (AAcid).

3. The method according to claim 1, wherein said concentration correlates to a severity of a condition of a human patient.

4. The method according to claim 3, wherein said condition is at least one of: a concussion, traumatic brain injury (TBI), and / or mild traumatic brain injury (mTBI);Alzheimer’s disease;Parkinson’s disease; cardiovascular disease; nonalcoholic fatty liver disease (NAFLD);cancer; diabetes; chronic obstructive pulmonary disease (COPD); kidney disease;Huntington’s disease (HD); amyotrophic lateral sclerosis (ALS); and bronchitis.

5. The method according to claim 1, further comprising the step of detecting a concentration of at least one other biomarker in said biological sample.

6. The method according to claim 5, wherein said at least one other biomarker and said aldehyde are simultaneously detected.

7. The method according to claim 1, wherein said evaluating step is performed using a portable electrochemistry device.

8. The method according to claim 7, further comprising displaying an indication relating to said concentration to a user of said device.

9. The method according to claim 1, wherein said evaluating step uses at least one of square wave voltammetry; chronoamperometry; and chronopotentiometry.

10. The method according to claim 1, wherein said biological sample is at least one of: blood; saliva; urine; sweat; and cerebrospinal fluid (CSF).

11. A system for detecting levels of a biomarker in a biological sample, said system comprising: a container for said biological sample and for a solution, said solution comprising a trapping agent selected to enhance an electrochemical response of said at least one biomarker in said biological sample;a contact-based sensor for sensing said electrochemical response when said biological sample is mixed with said solution in said container, said sensor being within said container and in contact with said mixture; a plurality of electrodes coupled to said sensor for applying one of a voltage and a current to said sensor and for receiving said electrochemical response through said sensor; and an electronics module for controlling said one of said voltage and said current and for measuring characteristics of said electrochemical response, wherein said biomarker is an aldehyde, wherein said trapping agent comprises at least one of an aniline and an aniline derivative , wherein said electronics module interfaces with a data processing device to thereby allow said data processing device to analyze said electrochemical response, and wherein said electronics module is coupled to said plurality of electrodes to allow for control of said one of said voltage and said current and for reception and measurement of said electrochemical response.

12. The system according to claim 11, wherein said trapping agent comprises at least one of 4-aminobiphenyl (4AB) and anthranilic acid (AAcid).

13. The system according to claim 11, wherein said concentration correlates to a severity of a condition of a human patient.

14. The system according to claim 11, wherein said condition is at least one of: a concussion, traumatic brain injury (TBI), and / or mild traumatic brain injury (mTBI);Alzheimer’s disease;Parkinson’s disease; cardiovascular disease; nonalcoholic fatty liver disease (NAFLD); cancer; diabetes; chronic obstructive pulmonary disease (COPD); kidney disease;Huntington’s disease (HD); amyotrophic lateral sclerosis (ALS); and bronchitis.

15. The system according to claim 11, wherein the system is configured to detect a concentration of at least one other biomarker in said biological sample.

16. The system according to claim 15, wherein said at least one other biomarker and said aldehyde are simultaneously detected.

17. The system according to claim 11, wherein said biological sample is at least one of: blood; saliva; urine; sweat; and cerebrospinal fluid (CSF).

18. The system according to claim 11, wherein said trapping agent is bound to a surface of at least one electrode of said plurality of electrodes.

19. A kit of parts for detecting levels of a biomarker in a biological sample, said kit of parts comprising: a container for said biological sample and for a solution, said solution comprising a trapping agent selected to enhance an electrochemical response of said at least one biomarker in said biological sample; a contact-based sensor for sensing said electrochemical response when said biological sample is mixed with said solution in said container, said sensor being for use within said container and in contact with said mixture;a plurality of electrodes coupled to said sensor for applying one of a voltage and a current to said sensor and for receiving said electrochemical response through said sensor; and an electronics module for controlling said one of said voltage and said current and for measuring characteristics of said electrochemical response, wherein said biomarker is an aldehyde, wherein said trapping agent comprises at least one of an aniline and an aniline derivative, wherein said electronics module interfaces with a data processing device to thereby allow said data processing device to analyze said electrochemical response, and wherein said electronics module is coupled to said plurality of electrodes to allow for control of said one of said voltage and said current and for reception and measurement of said electrochemical response.

20. The kit of parts according to claim 19, wherein said trapping agent comprises at least one of 4-aminobiphenyl (4AB) and anthranilic acid (AAcid).

21. The kit of parts according to claim 19, further comprising a collection apparatus for obtaining said biological sample.

22. The kit of parts according to claim 19, wherein said trapping agent is bound to a surface of at least one electrode of said plurality of electrodes.

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