Description:FIELD OF INVENTION
[0001] The present disclosure relates to electrochemical systems. More specifically, the present disclosure relates to an electrochemical biosensing device for detecting luteinizing hormone.
BACKGROUND
[0002] The detection of luteinizing hormone (LH) holds significance for monitoring ovulation, managing fertility, and identifying hormonal conditions such as polycystic ovary syndrome (PCOS). PCOS impacts a substantial number of women globally, highlighting the widespread relevance of accurate LH detection. Current methodologies for LH detection, however, exhibit several limitations that impede their utility for timely, convenient, and precise hormone monitoring. Understanding the dynamics of LH levels is vital for reproductive health and for addressing various endocrine system imbalances. The ability to accurately track LH surges can provide valuable information for family planning and for medical practitioners in assessing patient conditions. The ongoing development of more effective LH detection techniques remains a focus within medical and diagnostic fields due to the widespread need for such monitoring.
[0003] Laboratory-based techniques, such as enzyme-linked immunosorbent assay (ELISA), are recognized as a reliable standard for hormone quantification. While ELISA offers high accuracy in measuring hormone levels, it necessitates specialized laboratory infrastructure, trained personnel, and substantial processing times, often extending to several hours. Such requirements render ELISA unsuitable for rapid or point-of-care (PoC) applications, where quick results are often desired. Furthermore, the laboratory-based methods are often costly, which can limit their accessibility for frequent or home-based testing. The reliance on centralized facilities restricts the widespread application of ELISA for routine monitoring, particularly in settings where immediate feedback is beneficial.
[0004] Over-the-counter (OTC) ovulation prediction kits, frequently based on lateral flow immunoassays, provide a degree of convenience for users. However, these OTC kits are characterized by low sensitivity, subjective result interpretation, and semi-quantitative output. Such kits often fail to detect lower concentrations of LH, which can be problematic for women with irregular menstrual cycles or existing hormonal imbalances. The semi-quantitative nature of the results means that precise LH levels cannot be determined, and the visual assessment of results can lead to user misinterpretation. Moreover, the existing OTC methods are not designed for continuous or real-time monitoring, nor can they be readily integrated into wearable devices or broader digital health platforms, limiting their utility for comprehensive hormonal tracking.
[0005] A significant challenge in LH detection pertains to accessibility and affordability. Conventional LH detection methods, including ELISA, gas chromatography-mass spectrometry (GC-MS), proton transfer reaction-mass spectrometry (PTR-MS), and selected ion flow tube-mass spectrometry (SIFT-MS, are generally expensive, time-consuming, and demand centralized laboratory setups. The existing laboratory methods are frequently inaccessible to individuals residing in remote, low-resource, or underserved geographical regions. Such limitations restrict regular monitoring and can delay early identification of conditions like PCOS, thereby impacting timely medical intervention. The financial burden and logistical hurdles associated with these techniques present a barrier to equitable healthcare access for many individuals.
[0006] A notable absence exists in the domain of point-of-care (PoC) solutions for LH detection. The lack of real-time, portable, or wearable solutions prevents continuous hormone monitoring, which is particularly disadvantageous for women actively managing fertility or those contending with hormonal imbalances such as PCOS. The inability to conduct tests conveniently at home or in non-clinical settings restricts the frequency of monitoring and the ability to capture dynamic changes in LH levels, which are often indicative of important physiological events.
[0007] Therefore, there is a need to overcome the problems discussed above in luteinizing hormone detection.
OBJECTIVES
[0008] The primary objective of the present disclosure is to provide a flexible, and low-cost biosensor for real-time, point-of-care detection of luteinizing hormone for women's health applications.
[0009] Another objective of the present disclosure is to provide a biosensing device capable of rapid detection of luteinizing hormone.
[0010] Yet another objective of the present disclosure is to provide a biosensing device that is portable, and compatible with wearable and point-of-care applications, allowing for convenient monitoring.
[0011] Yet another objective of the present disclosure is to provide a biosensing device exhibiting high sensitivity and specificity for detection of luteinizing hormone.
[0012] Yet another objective of the present disclosure is to provide a biosensing device with long-term stability, capable of maintaining performance and repeatability under varying pH and storage conditions.
SUMMARY
[0013] The present disclosure addresses the limitations of existing approaches and provides a technical solution for rapid, sensitive, and point-of-care detection of luteinizing hormone. The present disclosure provides a biosensing device that overcomes the challenges associated with conventional laboratory-based techniques and over-the-counter test kits by offering a flexible and low-cost electrochemical biosensor platform interfaced with portable reader that enables real-time monitoring of luteinizing hormone levels. Enhanced sensitivity and specificity are achieved through the synergistic integration of a palladium-functionalized titanium carbide (Pd-Ti₃C₂) MXene nanocomposite and target-specific anti-LH antibodies, where the Pd-Ti₃C₂ matrix provides superior electron transfer and catalytic activity, while the immobilized antibodies confer high-affinity and selective binding toward luteinizing hormone (LH), collectively resulting in a highly efficient electrochemical transduction platform. The device provides rapid detection than traditional laboratory methods. The biosensor further demonstrates long-term stability and repeatability, with minimal degradation in performance, ensuring reliable results across various environmental conditions. The electrochemical biosensor platform is compatible with both non-invasive and invasive samples, increasing its versatility for different testing scenarios, and is adaptable for integration into or fabricates as wearable or handheld personal care devices, facilitating convenient and continuous hormone monitoring. The present disclosure addresses the need for accessible, accurate, and user-friendly luteinizing hormone detection, particularly beneficial for women's health applications such as fertility tracking and the management of conditions like Polycystic Ovary Syndrome (PCOS).
[0014] According to one aspect of the present disclosure, a biosensing device for sensitive and specific detection of luteinizing hormone (LH) is provided. The biosensing device comprises an electrochemical transduction platform comprising at least one electrode including a working electrode, a reference electrode, and a counter electrode; a conductive nanostructured interface deposited on the working electrode, the conductive nanostructured interface comprising a titanium carbide (Ti₃C₂Tx) MXene nanocomposite functionalized with metallic nanoparticles; and a biorecognition layer comprising monoclonal anti-luteinizing hormone (anti-LH) antibodies immobilized onto the conductive nanostructured interface via at least one immobilization mechanism selected from covalent coupling, non-covalent adsorption, cross-linking, or affinity-based interaction. The biosensing device is configured to detect and quantify luteinizing hormone upon exposure to a sample comprising luteinizing hormone, wherein the luteinizing hormone specifically binds to immobilized anti-LH antibodies on the working electrode, thereby modulating interfacial charge transfer kinetics and/or surface capacitance of the working electrode, and such alterations being transduced into a measurable electrochemical signal, wherein magnitude of the electrochemical signal is quantitatively proportional to concentration of luteinizing hormone in the sample. The sample is any one of non-invasive sample or an invasive sample.
[0015] The metallic nanoparticles are palladium (Pd) nanoparticles. The Pd nanoparticles are uniformly deposited onto a surface of Ti₃C₂ MXene, forming a palladium functionalized titanium carbide (Pd–Ti₃C₂) MXene nanocomposite that provides enhanced electrocatalytic activity, increased electroactive surface area, and improved antibody immobilization capability. The palladium nanoparticles are present in a form of nanodots, nanoclusters, or nanospheres.
[0016] The working electrode, the reference electrode, and the counter electrode are configured in a screen-printed, flexible, or miniaturized format. The working electrode is fabricated from carbon-based conductive materials selected from graphite, graphene, carbon nanotubes, and carbon black, thereby providing enhanced electrical conductivity, mechanical flexibility, and electrochemical stability for reliable detection of luteinizing hormone.
[0017] The biosensing device further comprises a portable electrochemical reader operatively interfaced with the electrochemical transduction platform. The portable electrochemical reader is configured to: apply a controlled electrochemical potential and/or current to the working electrode; acquire real-time electrochemical signals generated from biochemical interactions occurring at the surface of the working electrode; digitize acquired analog signals via an onboard analog-to-digital converter (ADC); process digitized signals through embedded firmware and/or software algorithms for baseline correction, noise filtering, and calibration; and determine quantitative levels of luteinizing hormone in the sample by correlating processed electrochemical signals with stored calibration models. The portable electrochemical reader optionally comprises a microcontroller, a rechargeable power source, a display unit, and/or a wireless communication module for enabling real-time, user-friendly, point-of-care detection of luteinizing hormone.
[0018] The biosensing device is characterized by a linear detection range of 1–200 milli-international units per milliliter (mIU/mL) of luteinizing hormone and exhibits a limit of detection (LoD) of about 0.03 mIU/mL or lower, thereby enabling highly sensitive, accurate, and reproducible quantification of luteinizing hormone across clinically relevant concentrations.
[0019] The biosensing device is configured to be integrated into or fabricated as wearable or handheld personal care devices selected from wristbands, smart patches, smart watches, handheld diagnostic readers, or smartphone-integrated accessories, thereby enabling real-time, continuous, and user-friendly monitoring of luteinizing hormone levels in biological fluids, for applications in personal healthcare, fertility tracking, and point-of-care diagnostics.
[0020] The biosensing device exhibits high specificity toward luteinizing hormone (LH) with negligible cross-reactivity to potential interfering analytes including thyroid stimulating hormone (TSH), immunoglobulin G (IgG), uric acid (UA), and ascorbic acid (AA).
[0021] The biosensing device demonstrates reproducibility with a relative standard deviation (RSD) of less than 5% and maintains long-term stability with a shelf life exceeding 50 days, exhibiting no more than about 3.2% degradation in electrochemical current response under optimized conditions.
[0022] The biosensing device is configured to detect and quantify the luteinizing hormone within 30 seconds of sample introduction, as evidenced by rapid electrochemical signal generation in cyclic voltammetry profiles across a dynamic concentration range (1–200 mIU/mL).
[0023] According to another aspect of the present disclosure, a method of producing a biosensing device for sensitive and specific detection of luteinizing hormone (LH) is provided. The method comprises providing an electrochemical platform comprising at least one electrode including a working electrode, a reference electrode, and a counter electrode; depositing a conductive nanostructured interface comprising a titanium carbide (Ti₃C₂Tx) MXene nanocomposite functionalized with metallic nanoparticles; and immobilizing a biorecognition layer comprising monoclonal anti-luteinizing hormone (anti-LH) antibodies onto conductive nanostructured interface via at least one immobilization mechanism selected from covalent coupling, non-covalent adsorption, cross-linking, or affinity-based interaction. The metallic nanoparticles are palladium nanoparticles, wherein the Pd nanoparticles are uniformly deposited onto a surface of Ti₃C₂ MXene, forming a palladium functionalized titanium carbide (Pd–Ti₃C₂) MXene nanocomposite. The palladium nanoparticles are present in a form of nanodots, nanoclusters, or nanospheres.
[0024] The method further comprises depositing the Pd nanoparticles uniformly onto a surface of Ti₃C₂ MXene, forming a palladium functionalized titanium carbide (Pd–Ti₃C₂) MXene nanocomposite on the surface of the working electrode, thereby providing enhanced electrocatalytic activity, increased electroactive surface area, and improved immobilization efficiency of anti-luteinizing hormone antibodies for highly sensitive detection. The deposition is carried out by drop-casting of pre-synthesized nanoparticles.
[0025] The method further comprises interfacing a portable electrochemical reader with the electrochemical transduction platform. The portable electrochemical is configured for apply a controlled electrochemical potential and/or current to the working electrode; acquire real-time electrochemical signals generated from biochemical interactions occurring at the surface of the working electrode; digitize acquired analog signals via an onboard analog-to-digital converter (ADC); process digitized signals through embedded firmware and/or software algorithms for baseline correction, noise filtering, and calibration; and determine quantitative levels of luteinizing hormone in the sample by correlating processed electrochemical signals with stored calibration models. The portable electrochemical reader optionally comprises a microcontroller, a rechargeable power source, a display unit, and/or a wireless communication module for enabling real-time, user-friendly, point-of-care detection of luteinizing hormone.
[0026] The method further comprises integrating or fabricating the biosensing device as wearable or handheld personal care devices selected from wristbands, smart patches, smart watches, handheld diagnostic readers, or smartphone-integrated accessories.
[0027] The foregoing paragraphs have been provided by way of general introduction and are not intended to limit the scope of the following claims. The described embodiments, together with further advantages, will be best understood by reference to the following detailed description taken in conjunction with the accompanying drawings.
BRIEF DESCRIPTION OF DRAWINGS
[0028] FIG. 1 is a schematic illustration of a biosensing device in accordance with the present disclosure.
[0029] FIG. 2 is a schematic illustration of an electrochemical transduction platform in accordance with the present disclosure.
[0030] FIG. 3 is a schematic illustration of a working electrode of an electrochemical transduction platform in accordance with the present disclosure.
[0031] FIG. 4 is a block diagram illustrating a portable electrochemical reader in accordance with the present disclosure.
[0032] FIG. 5a is a graphical representation illustrating cyclic voltammograms (CVs) of a biosensing device at different luteinizing hormone (LH) concentrations (1–200 mIU/mL) in accordance with the present disclosure.
[0033] FIG. 5b is a graphical representation of a calibration plot illustrating the linear relationship between peak current and LH concentration, further indicating the limit of detection (LoD = 0.03 mIU/mL) in accordance with the present disclosure.
[0034] FIG. 5c is a graphical representation illustrating resistance versus LH concentration curves obtained using a lab-based measurement system.
[0035] FIG. 5d is a graphical representation illustrating resistance as a function of LH concentration, measured using a biosensing device in accordance with the present disclosure.
[0036] FIG. 6a is a graphical representation of a calibration plot depicting current response as a function of LH concentration (1–200 mIU/mL) measured in both ferro/ferricyanide electrolyte and human serum in accordance with the present disclosure.
[0037] FIG. 6b is a bar graph illustrating peak current values for varying LH concentrations in ferro/ferricyanide system versus human serum in accordance with the present disclosure.
[0038] FIG. 7a is a bar graph illustrating sensor's current responses to various analytes in accordance with the present disclosure.
[0039] FIG. 7b is a bar graph illustrating reproducibility of an electrochemical transduction platform in accordance with the present disclosure.
[0040] FIG. 7c is a bar graph illustrating long-term stability of an electrochemical transduction platform in accordance with the present disclosure.
[0041] FIG. 8 is a graphical representation illustrating cyclic voltammograms (CVs) of a biosensing device recorded at fixed antibody of luteinizing hormone (Anti-LH) concentrations with varying time from (0-60 sec) in accordance with the present disclosure.
[0042] FIG. 9a is a graphical representation illustrating cyclic voltammograms of bare CSPE, CSPE/ Pd-Ti3C2Tx, and CSPE/ Pd-Ti3C2Tx /Anti-LH in accordance with the present disclosure.
[0043] FIG. 9b is a graphical representation of differential pulse voltammetry (DPV) obtained for the same electrodes, illustrating the changes in peak current observed after each successive modification in accordance with the present disclosure.
[0044] FIG. 9c is a graphical representation illustrating electrochemical impedance spectroscopy (EIS) plots for bare CSPE, CSPE/Pd-Ti₃C₂Tx, and CSPE/Pd-Ti₃C₂Tx/Anti-LH, demonstrating the variation in charge transfer resistance corresponding to each stage of electrode modification in accordance with the present disclosure.
[0045] FIG. 9d is a graphical representation illustrating cyclic voltammograms (CVs) of CSPE/Pd-Ti₃C₂Tx/Anti-LH recorded at varying scan rates (10–100 mV/s), depicting the dependence of current response on the applied scan rate in accordance with the present disclosure.
[0046] FIG. 9e is a graphical representation illustrating anodic and cathodic peak currents (Ipa and Ipc) as a function of the square root of the scan rate, confirming the electrochemical behavior of the modified electrode in accordance with the present disclosure.
[0047] FIG. 10 is a flowchart illustrating a method of producing an electrochemical biosensing device in accordance with the present disclosure.
DETAILED DESCRIPTION OF THE PRESENT DISCLOSURE
[0048] Aspects of the present invention are best understood by reference to the description set forth herein. All the aspects described herein will be better appreciated and understood when considered in conjunction with the following descriptions. It should be understood, however, that the following descriptions, while indicating preferred aspects and numerous specific details thereof, are given by way of illustration only and should not be treated as limitations. Changes and modifications may be made within the scope herein without departing from the spirit and scope thereof, and the present invention herein includes all such modifications.
[0049] As mentioned above, there is a need for a technical solution to solve aforementioned technical problems in detection of luteinizing hormone (LH). The present invention provides a biosensing device that utilizes a flexible carbon screen-printed electrode as its base, upon which a conductive nanostructured interface comprising a palladium-functionalized titanium carbide (Pd-Ti3C2) MXene nanocomposite is deposited. The nanocomposite significantly enhances electron transfer and catalytic activity, leading to improved sensitivity and signal amplification. The working electrode is further functionalized with a biorecognition layer of monoclonal anti-luteinizing hormone antibodies. The layered structure of the electrochemical platform enables specific and sensitive detection of LH through modulation of interfacial charge transfer kinetics and/or surface capacitance upon binding of LH to the antibodies. The biosensing device offers rapid detection of LH, a wide linear range of 1-200 milli-international units per milliliter (mIU/mL), and low detection and quantification limits. Further, integration of the electrochemical platform with a portable electrochemical reader enhances the functionality and user-friendliness of the biosensing device. The portable reader is configured to interface seamlessly with the electrochemical transduction platform, allowing for the application of controlled electrochemical potentials or currents to the working electrode, thereby obtaining real-time electrochemical signals generated as a result of biochemical interactions occurring at the surface of the working electrode. The portable reader then analyses the signal output corresponding to luteinizing hormone concentration. Such integration enables rapid and automated quantification of LH levels, providing users with easily interpretable results within seconds. The portable nature of the reader, combined with its data processing capabilities, facilitates on-the-spot analysis in various settings, from clinical environments to home use. The combination of the biosensor and portable reader represents a comprehensive solution for point-of-care LH monitoring, offering the accuracy of laboratory-grade equipment in a compact, user-friendly format suitable for widespread adoption in personal health management and clinical diagnostics.
[0050] Now referring to FIGS. 1-2, FIG. 1 is a schematic diagram illustrating an electrochemical biosensing device 100 in accordance with the present disclosure. FIG. 2 is a schematic illustration of the electrochemical transduction platform 102 in accordance with the present disclosure. The biosensing device 100 comprises an electrochemical transduction platform 102 and a portable electrochemical reader 104. The electrochemical transduction platform 102 includes a reference electrode 106, a counter electrode 108, and a working electrode 110 (seen in FIG. 2). The electrochemical transduction platform 102 is configured to detect luteinizing hormone (LH) from a single drop of biological sample. In some embodiments, the biological sample is any of blood, serum, plasma, saliva, urine, sweat, or tear fluid. The electrochemical transduction platform 102 may be adapted to accommodate variations in sample matrix composition and viscosity, and to selectively capture LH through immobilized antibodies, aptamers, or molecularly imprinted polymers on the electrode surface. The electrochemical transduction platform 102 refers to a sensing interface that converts a biochemical interaction (such as enzyme–substrate binding, antigen–antibody recognition, or analyte adsorption) into a measurable electrical signal, typically involving current, voltage, or impedance changes. The working electrode 110 refers to a sensing element configured to selectively interact with target analytes and generate electrochemical signals proportional to its concentration. The reference electrode 106 refers to a stable electrode with a well-defined and constant potential that provides a fixed reference point for measuring the potential changes at the working electrode 110 during electrochemical measurements. The counter electrode 108 refers to an electrode that completes the electrochemical circuit by allowing current to flow between itself and the working electrode(s), typically made from an inert conductive material to ensure minimal interference with the sensing process.
[0051] The electrochemical transduction platform 102 comprises a substrate that provides mechanical support upon which a set of electrodes including the working electrode 110, the reference electrode 106, and the counter electrode 108 are patterned in a screen-printed, flexible, or miniaturized configuration. In some embodiments, the working electrode 110 is fabricated from carbon-based conductive materials such as graphite, graphene, carbon nanotubes, or carbon black, selected to impart superior electrical conductivity, mechanical flexibility, and electrochemical stability.
[0052] In some embodiments, the reference electrode 106 is fabricated using a material selected from a group comprising silver/silver chloride (Ag/AgCl), mercury/mercury chloride (Hg/Hg2Cl2), copper/copper sulfate (Cu/CuSO4), platinum (Pt), gold (Au), iridium oxide (IrO2), palladium/hydrogen (Pd/H2), printed carbon, conductive polymers doped with specific ions, and zinc/zinc sulfate (Zn/ZnSO4). The reference electrode 106 ensures that the potential applied to or measured from the working electrode 110 is accurately controlled and interpreted. The counter electrode 108 completes the electrical circuit and balances the current generated at the working electrode 110. In some embodiments, the counter electrode 108 is made of an inert conductive material such as platinum or carbon, chosen for its ability to support the necessary electrochemical reactions without interfering with the sensing processes.
[0053] As seen in FIG. 3, the surface of the working electrode 110 is further modified with nanoparticle-functionalized titanium carbide MXene nanocomposite, thereby forming a conductive nanostructured interface. The nanoparticle is palladium nanoparticle. The Pd nanoparticles are uniformly deposited onto a surface of Ti₃C₂ MXene, forming a palladium functionalized titanium carbide (Pd–Ti₃C₂) MXene nanocomposite. The modification of the surface of the working electrode 110 enhances the electrochemical performance and functionality of the working electrode 110. The palladium nanoparticles may be in the form of nanodots, nanoclusters, or nanospheres, and deposited by a process of drop casting, thereby providing enhanced electrocatalytic activity, increased electroactive surface area, and improved immobilization efficiency of anti-luteinizing hormone antibodies for highly sensitive detection.
[0054] The working electrode 110 is further functionalized with monoclonal anti-luteinizing hormone (anti-LH) antibodies, thereby enabling the immunosensing of luteinizing hormone on the MXene-modified electrode surface of the working electrode 110. The monoclonal anti-luteinizing hormone (anti-LH) antibodies are immobilized onto the nanoparticle-functionalized MXene nanocomposite modified surface of the working electrode through at least one immobilization mechanism selected from covalent coupling, non-covalent adsorption, cross-linking, or affinity-based interaction.
[0055] The portable electrochemical reader 104 interfaces with the electrochemical transduction platform 102 to record and analyze the electrochemical responses from the working electrode 110. The portable electrochemical reader 104 is configured to apply a controlled electrochemical potential or current to the working electrode 110; obtaining real-time electrochemical signals generated as a result of biochemical interactions occurring at a surface of the working electrode 110; digitizing acquired analog signals via an onboard analog-to-digital converter (ADC); processing the digitized signals through embedded firmware and/or software algorithms for baseline correction, noise filtering, and calibration; and determining quantitative levels of luteinizing hormone in the sample by executing embedded algorithms or firmware to analyze signal output corresponding to luteinizing hormone concentration.
[0056] The electrochemical biosensing device 100 operates by applying a single drop of biological fluid onto the electrochemical transduction platform 102. The LH present in the biological fluid binds to the anti-LH antibodies immobilized on the working electrode 110. The binding event alters the electrode's interfacial properties, which is detected through changes in impedance or voltammetric response.
[0057] In some embodiments, the biosensing device 100 is configured to be integrated into or fabricated as wearable or handheld personal care devices selected from wristbands, smart patches, smart watches, handheld diagnostic readers, or smartphone-integrated accessories, thereby enabling real-time, continuous, and user-friendly monitoring of luteinizing hormone levels in biological fluids, for applications in personal healthcare, fertility tracking, and point-of-care diagnostics. The diverse form factors enable versatile application of the biosensing technology across different user preferences and lifestyle needs. For instance, integration into smartwatches or fitness trackers allows for continuous, non-intrusive monitoring throughout the day, while adhesive patches offer discreet, longer-term wear for extended analysis periods. Smart contact lenses could provide a unique approach to hormone monitoring through tear analysis, offering a non-invasive method for frequent measurements. Smartphone accessories, such as attachable modules or cases with integrated biosensors, leverage the ubiquity of smartphones to make LH monitoring more accessible and user-friendly. Portable urinalysis devices equipped with the biosensing apparatus can offer convenient at-home testing with rapid results, particularly useful for ovulation prediction and fertility tracking. The integration of the biosensing apparatus into these various form factors not only enhances the convenience and regularity of LH monitoring but also opens up new possibilities for comprehensive women's health management in everyday life.
[0058] The biosensing device 100 demonstrates analytical performance in the detection and quantification of luteinizing hormone (LH). The device 100 provides a wide linear detection range spanning from 1 to 200 milli-international units per milliliter (mIU/mL), encompassing the clinically relevant concentrations of LH in various physiological states. The broad range enables accurate measurement of LH levels from basal concentrations to surge levels during ovulation. The device 100 exhibits a remarkably low limit of detection (LoD) of 3 mIU/mL, allowing for the identification of minute quantities of LH in biological samples. The high sensitivity is crucial for detecting subtle hormonal changes and early-stage LH surges. Furthermore, the electrochemical transduction platform 102 boasts a limit of quantification (LoQ) of 8 mIU/mL, ensuring reliable and precise quantitative measurements even at low hormone concentrations. Such performance metrics are achieved through the synergistic combination of the Pd-Ti₃C₂ MXene nanocomposite interface and the highly specific anti-LH antibodies, resulting in enhanced signal amplification and reduced background noise. The apparatus 100 maintains high level of performance across various sample matrices, including serum, urine, and saliva, demonstrating its versatility in different testing scenarios. Accordingly, the biosensing device 100 accurately performs hormone monitoring in applications ranging from fertility tracking to the diagnosis and management of hormonal disorders.
[0059] Further, the biosensing device 100 achieves rapid detection, providing results within 30 seconds, which significantly enhances its utility for point-of-care applications and real-time monitoring. The device 100 demonstrates long-term stability, maintaining its performance for at least 1.5 months with minimal degradation, exhibiting no more than 3.2% reduction in current response over this period. The stability extends across varying pH levels and storage conditions, ensuring consistent and repeatable results in diverse testing environments. The combination of rapid detection, high sensitivity, and long-term stability is achieved through the Pd-Ti₃C₂ MXene nanocomposite interface and highly specific anti-LH antibodies, resulting in a robust and versatile biosensing platform 100.
[0060] The device 100 exhibits high specificity toward luteinizing hormone (LH) with negligible cross-reactivity to potential interfering analytes including thyroid stimulating hormone (TSH), immunoglobulin G (IgG), uric acid (UA), and ascorbic acid (AA), demonstrates reproducibility with a relative standard deviation (RSD) of less than 5% across at least seven independently fabricated electrodes, and maintains long-term stability with a shelf life exceeding 50 days, exhibiting no more than about 3.2% degradation in electrochemical current response under optimized conditions.
[0061] With reference to FIGS 1-2, FIG. 3 is a schematic illustration of the working electrode 110 in accordance with the present disclosure. FIG. 3 depicts the layered structure of the working electrode 110 and the immunoaffinity-based detection process for luteinizing hormone. The working electrode 110 comprises a carbon layer 302, a conductive nanostructured interface 304, and an immunoaffinity-based biorecognition layer 306.
[0062] The carbon layer 302 forms the base of the working electrode 110. The carbon layer 302 is typically composed of screen-printed carbon ink, which offers excellent electrical conductivity and electrochemical stability. The carbon layer 302 provides a cost-effective and versatile substrate for further modification and functionalization. Alternative materials for the carbon layer could include graphite, graphene, carbon nanotubes, carbon black, each offering unique properties that may enhance the overall performance of the electrode.
[0063] The conductive nanostructured interface 304 is deposited on top of the carbon layer 302. The conductive nanostructured interface 304 comprises a Pd-Ti₃C₂ MXene nanocomposite, which is synthesized through a two-step process. First, Ti₃C₂ MXene is prepared via selective etching of aluminum layers from Ti₃AlC₂ using a chemical reduction method, typically involving hydrofluoric acid (HF) as the etching agent. The etching process removes the aluminum layers from the Ti₃AlC₂ precursor, resulting in the formation of two-dimensional Ti₃C₂ MXene sheets with high surface area and excellent electrical conductivity. Subsequently, palladium (Pd) nanoparticles are uniformly deposited onto the surface of the prepared Ti₃C₂ MXene, a crucial step for enhancing the electrical conductivity and catalytic properties of the nanocomposite. The uniform deposition of Pd nanoparticles creates numerous active sites on the MXene surface, significantly improving the electron transfer capabilities and the catalytic activity of the interface 304. The Pd nanoparticles are deposited by a process of drop casting, where a predetermined volume of the nanoparticle suspension is carefully dispensed onto the prepared Ti₃C₂ MXene, followed by drying under controlled conditions to allow solvent evaporation and uniform adhesion of the nanoparticles onto the electrode substrate. The process may be repeated multiple times to achieve the desired nanoparticle loading and surface coverage. The resulting Pd-Ti₃C₂ MXene nanocomposite combines the advantages of both materials such as the high surface area and conductivity of MXene with the catalytic properties of palladium nanoparticles. The synergistic combination creates a highly effective conductive nanostructured interface 304 that significantly enhances the sensitivity and performance of the biosensing apparatus 100, enabling rapid and efficient electron transfer during the electrochemical detection of luteinizing hormone.
[0064] The immunoaffinity-based biorecognition layer 306 is formed on the conductive nanostructured interface 304 of the working electrode 110. The immunoaffinity-based detection element consists of anti-LH antibodies specific for LH 308 are immobilized on the conductive nanostructured interface 304. The immobilization process is carried out using covalent coupling, non-covalent adsorption, cross-linking, or affinity-based interaction, ensuring stable attachment of the antibodies to the Pd-Ti₃C₂ MXene nanocomposite surface. Covalent bonding may involve the use of cross-linking agents to form stable chemical bonds between the antibodies and the nanocomposite surface, while non-covalent immobilization can utilize methods such as physical adsorption or bioaffinity interactions. The orientation of the immobilized antibodies is optimized to ensure maximum accessibility of their antigen-binding sites, thereby enhancing the capture efficiency of LH molecules. The biorecognition layer 306 selectively recognizes and binds with LH 308 from biological samples. The integration of the highly specific biorecognition element with the conductive nanostructured interface 304 creates a sensing platform capable of detecting minute concentrations of LH with high accuracy and reliability.
[0065] The LH 308 represents the target analyte in the sample. When a drop of serum is applied to the working electrode 110, LH molecules present in the sample specifically bind to the immobilized anti-LH antibodies on the working electrode 110. The antigen-antibody interaction forms a complex layer on the electrode surface of the working electrode 110, altering the interfacial electron transfer properties. The change in electron transfer characteristics is then measured using electrochemical techniques such as electrochemical impedance spectroscopy (EIS) or differential pulse voltammetry (DPV).
[0066] The immunosensing approach allows for label-free, real-time detection of LH from biological sample. The use of Pd-Ti₃C₂ nanocomposite as the nanostructured interface enhances the sensitivity and stability of the immunosensor.
[0067] In some embodiments, the sample is selected from biological fluids, including but not limited to whole blood, serum, plasma, saliva, and urine. In certain aspects, serum samples may be derived from patient blood, processed to remove cellular components, while plasma samples may be collected using anticoagulants to preserve clotting factors. Saliva and urine offer non-invasive or minimally invasive alternatives for sample acquisition, thereby reducing patient discomfort and increasing user compliance. The ability of the biosensor to function reliably across these varied biological matrices enables versatile detection of luteinizing hormone for use in both clinical laboratory settings and portable, point-of-care diagnostic platforms.
[0068] FIG. 4 is a block diagram illustrating the portable electrochemical reader 104 in accordance with the present disclosure. The portable electrochemical reader 104 includes, but not limited to, a potentiostat circuit 402 for controlling and measuring electrode potentials and currents; an analog-to-digital converter (ADC) 404 for signal digitization; a microcontroller 406 for overall operation management; dedicated modules for electrochemical techniques such as amperometry, differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS); signal processing units 408 for filtering and amplifying acquired signals; memory 410 for data and parameter storage; a rechargeable power source 412 for regulating power supply; a user interface 414 with display and input options; communication modules 416 for data transfer to external devices; and data analysis software for interpreting raw data into meaningful LH readings. The portable electrochemical reader 104 also incorporates calibration and quality control features to ensure measurement accuracy. The combination of these components enhances the overall usability and effectiveness of the biosensing apparatus, making it suitable for various settings from clinical environments to point-of-care diagnostics and field testing.
[0069] FIG. 5a is a graphical representation illustrating cyclic voltammograms (CVs) of a biosensing device 100 at different luteinizing hormone (LH) concentrations (1–200 mIU/mL) in accordance with the present disclosure. Each successive curve demonstrates a distinct change in current response as the LH concentration increases, thereby confirming the electrochemical activity of the modified working electrode surface and its ability to quantitatively detect LH across a broad concentration range. The inset portion of FIG. 5a depicts the current responses obtained at low LH concentrations, thereby highlighting the sensitivity of the biosensing device in detecting trace amounts of the analyte
[0070] FIG. 5b is a graphical representation of a calibration plot illustrating the linear relationship between peak current and LH concentration, further indicating the limit of detection (LoD = 0.03 mIU/mL) in accordance with the present disclosure. The black points and line represent the experimentally measured current values at different LH concentrations, while the red line corresponds to the linear regression fit of the data, with an R² value of 0.998. The linearity validates the reproducibility and reliability of the device, while the limit of detection (LoD = 0.03 mIU/mL) highlights its capability to detect trace levels of LH with high sensitivity.
[0071] FIG. 5c is a graphical representation illustrating resistance versus LH concentration curves obtained using a lab-based measurement system. FIG. 5d is a graphical representation illustrating resistance as a function of LH concentration, measured using the biosensing device 100 in accordance with the present disclosure. The black points and line represent the resistance values experimentally measured by the biosensing device 100, while the red line corresponds to the regression fit (R² = 0.998), showing excellent agreement. The data exhibit a similar decreasing resistance trend with increasing LH concentration as observed in the laboratory setup, thereby demonstrating that the biosensing device 100 can reliably operate not only under laboratory conditions but also in a portable, point-of-care format for real-world diagnostic applications.
[0072] FIG. 6a is a graphical representation of a calibration plot depicting current response as a function of LH concentration (1–200 mIU/mL) measured in both ferro/ferricyanide electrolyte and human serum in accordance with the present disclosure. The graphical representation includes two sets of data: measurements performed in a standard ferro/ferricyanide electrolyte system, while the measurements performed in human serum samples. Both datasets exhibit a positive correlation between current response and LH concentration, with minimal deviation between the ferro/ferricyanide and serum matrices. This demonstrates that the biosensor maintains accuracy and reliability when transitioning from a model electrolyte system to complex biological fluids. The close overlap of the data further confirms the robustness of the biorecognition interface and its suitability for practical diagnostic applications.
[0073] FIG. 6b is a bar graph illustrating peak current values for varying LH concentrations in ferro/ferricyanide system versus human serum in accordance with the present disclosure. The bar plots show a consistent increase in peak current with increasing LH concentration in both systems, with slightly lower absolute current responses in serum due to the presence of biological interferents. Nevertheless, the trend remains highly proportional across the tested concentration range, thereby confirming the sensor’s ability to deliver reliable quantification of LH in real clinical samples.
[0074] FIG. 7a is a bar graph illustrating sensor's current responses to various analytes in accordance with the present disclosure. The bar labeled LH (luteinizing hormone) shows a significantly higher current response (49.7 µA) compared to potential interferents such as thyroid-stimulating hormone (TSH), albumin, ascorbic acid, uric acid, glucose, and serum components, which exhibit much lower signals ranging from 5.3 µA to 16.4 µA. The “No Target” control also shows a negligible response. This confirms the sensor’s high specificity toward LH, with minimal cross-reactivity against non-target biomolecules typically present in biological samples.
[0075] FIG. 7b is a bar graph illustrating reproducibility of an electrochemical transduction platform in accordance with the present disclosure. The recorded current responses remain highly consistent, with values ranging from 48.9 µA to 49.7 µA, and only minor variation observed. The low relative standard deviation (RSD) underscores the reproducibility and robustness of the fabrication process, ensuring reliability of the biosensor for practical applications.
[0076] FIG. 7c is a bar graph illustrating long-term stability of an electrochemical transduction platform in accordance with the present disclosure. The electrochemical transduction platform maintained a nearly constant current response throughout the testing period, with only a minor degradation of 3.32% observed at day 50 compared to the initial value. This result demonstrates excellent storage stability of the biosensor, ensuring extended shelf life and reliability for real-world use in diagnostic settings.
[0077] FIG. 8 is a graphical representation illustrating cyclic voltammograms (CVs) of a biosensing device recorded at fixed antibody of luteinizing hormone (Anti-LH) concentrations with varying time from (0-60 sec) in accordance with the present disclosure. Each CV curve corresponds to a specific incubation time interval. As shown, the current response increases progressively with time, reflecting enhanced antibody–antigen interactions at the electrode surface. Importantly, a distinct and reliable electrochemical signal is achieved within the first 30 seconds, thereby enabling rapid detection of luteinizing hormone as claimed in the present disclosure. While extended incubation up to 60 seconds yields a marginally higher current response, such additional time is not essential for effective analyte detection. The inset highlights the current responses in the low-potential region, further confirming the rapid and time-dependent binding kinetics of the biosensor.
[0078] FIG. 9a is a graphical representation illustrating cyclic voltammograms of bare CSPE, CSPE/ Pd-Ti3C2Tx, and CSPE/ Pd-Ti3C2Tx /Anti-LH in accordance with the present disclosure. The bare CSPE shows relatively low current response, while modification with Pd-Ti₃C₂Tx significantly enhances the current due to improved electron transfer properties. Upon immobilization of Anti-LH, the current decreases, indicating successful surface functionalization and antibody binding, which partially hinders electron transfer.
[0079] FIG. 9b is a graphical representation of differential pulse voltammetry (DPV) obtained for the same electrodes, illustrating the changes in peak current observed after each successive modification in accordance with the present disclosure. A clear variation in peak current is observed after each successive modification: the Pd-Ti₃C₂Tx-modified electrode exhibits higher current compared to bare CSPE, while the Anti-LH modification leads to a reduction in peak current. This sequential change in current response validates each stage of electrode assembly and confirms the effective immobilization of the biorecognition element.
[0080] FIG. 9c is a graphical representation illustrating electrochemical impedance spectroscopy (EIS) plots for bare CSPE, CSPE/Pd-Ti₃C₂Tx, and CSPE/Pd-Ti₃C₂Tx/Anti-LH, demonstrating the variation in charge transfer resistance corresponding to each stage of electrode modification in accordance with the present disclosure. The bare CSPE displays high charge transfer resistance, which decreases markedly upon Pd-Ti₃C₂Tx modification due to the excellent conductivity of the nanomaterial. Following Anti-LH immobilization, the charge transfer resistance increases again, reflecting the insulating nature of the biomolecular layer. These results provide additional evidence of successful stepwise electrode modification.
[0081] FIG. 9d is a graphical representation illustrating cyclic voltammograms (CVs) of CSPE/Pd-Ti₃C₂Tx/Anti-LH recorded at varying scan rates (10–100 mV/s), depicting the dependence of current response on the applied scan rate in accordance with the present disclosure. The observed increase in peak current with increasing scan rate demonstrates the electrode’s electrochemical responsiveness and supports diffusion-controlled electron transfer kinetics at the electrode–electrolyte interface.
[0082] FIG. 9e is a graphical representation illustrating anodic and cathodic peak currents (Ipa and Ipc) as a function of the square root of the scan rate, confirming the electrochemical behavior of the modified electrode in accordance with the present disclosure. The linear dependence observed for both Ipa and Ipc, with correlation coefficients close to unity (R² ≈ 0.998), confirms that the redox processes at the modified electrode surface are diffusion-controlled and that the electrode exhibits stable electrochemical behavior suitable for biosensing applications.
[0083] With reference to FIGS. 1-9e, FIG. 10 is a flowchart illustrating a method of producing an electrochemical biosensing device 100 and detecting LH using the electrochemical biosensing device 100 in accordance with the present disclosure. At step 1002, the method includes providing the screen-printed electrochemical transduction platform 102 comprising the working electrode 110, the reference electrode 106, and the counter electrode 108. In some embodiments, the preparation of the platform 102 involves selecting and cleaning a substrate material, for instance polyethylene terephthalate (PET) or ceramic; designing the working electrode 110, the reference electrode 106, and the counter electrode 108; printing carbon ink onto the substrate in a single step, forming the base layer for all working electrodes; drying and curing the printed carbon layer under controlled temperature conditions to ensure proper adhesion and conductivity; printing the reference electrode material, for instance silver/silver chloride ink, onto a designated area; if required, printing an additional layer of conductive material (e.g., platinum-based ink) over the carbon layer in a counter electrode area to improve its performance; printing an insulation layer over the electrode traces, leaving only the active areas of the electrodes and contact pads exposed. performing final curing to ensure the stability and durability of all printed layers.
[0084] At step 1004, the method includes depositing a conductive nanostructured interface 304 comprising Pd nanoparticles functionalized titanium carbide MXene nanocomposite on the working electrode 110. The deposition process begins with the synthesis of Ti₃C₂ MXene which is prepared via selective etching of aluminum layers from Ti₃AlC₂ using a chemical reduction method, typically involving hydrofluoric acid (HF) as the etching agent. The etching process removes the aluminum layers from the Ti₃AlC₂ precursor, resulting in the formation of two-dimensional Ti₃C₂ MXene sheets with high surface area and excellent electrical conductivity. Subsequently, palladium (Pd) nanoparticles are uniformly deposited, using a process of drop casting, onto the surface of the prepared Ti₃C₂ MXene, a crucial step for enhancing the electrical conductivity and catalytic properties of the nanocomposite. The uniform deposition of Pd nanoparticles creates numerous active sites on the MXene surface, significantly improving the electron transfer capabilities and the catalytic activity of the interface 304. The resulting Pd-Ti₃C₂ MXene nanocomposite combines the advantages of both materials such as the high surface area and conductivity of MXene with the catalytic properties of palladium nanoparticles. The synergistic combination creates a highly effective conductive nanostructured interface 304 that significantly enhances the sensitivity and performance of the biosensing apparatus 100, enabling rapid and efficient electron transfer during the electrochemical detection of luteinizing hormone.
[0085] At step 1006, the method includes immobilizing a biorecognition layer 306 comprising monoclonal anti-luteinizing hormone (anti-LH) antibodies onto a nanoparticle-functionalized titanium carbide MXene nanocomposite modified surface of the working electrode 110 via at least one immobilization mechanism selected from covalent coupling, non-covalent adsorption, cross-linking, or affinity-based interaction. The immobilization step involves selecting high-affinity monoclonal anti-LH antibodies, chosen for their specificity and sensitivity to luteinizing hormone. The immobilization process is optimized to ensure maximum retention of antibody activity and optimal orientation for efficient LH capture. For covalent bonding, techniques such as carbodiimide chemistry or click chemistry may be employed, forming stable amide or triazole linkages between the antibodies and the nanocomposite surface. Non-covalent immobilization strategies might include biotin-streptavidin interactions, electrostatic adsorption, or hydrophobic interactions, each offering unique advantages in terms of simplicity and preserved antibody conformation. The density of immobilized antibodies is controlled to prevent overcrowding and steric hindrance, ensuring optimal binding kinetics and sensor performance. The synergy between the nanostructured Pd-Ti₃C₂ MXene interface and the immobilized anti-LH antibodies results in the biorecognition layer 306 with enhanced stability, sensitivity, and specificity, capable of rapid and accurate LH detection across a wide concentration range.
[0086] At step 1008, the method includes applying the biological sample to the electrode platform 102. For instance, a small volume (e.g., 5-50 μL) of human serum or whole blood, is applied onto the working electrode surface. Techniques such as micropipetting, automated dispensing systems, or specialized microfluidic channels may be employed to achieve accurate and reproducible sample delivery. The sample volume is optimized to provide sufficient analyte for both sensing mechanisms while minimizing the required blood draw from the patient. Upon application, the sample rapidly spreads across the electrode surface. This spreading ensures efficient contact between the analytes (LH) and the respective biorecognition elements (anti-LH antibodies).
[0087] At step 1010, the method includes detecting and quantifying luteinizing hormone in a sample by facilitating binding of luteinizing hormone with immobilized anti-LH antibodies, thereby modulating interfacial charge transfer kinetics and/or surface capacitance of the working electrode 110, and generating electrochemical signal proportional to concentration of luteinizing hormone, in response to changes in the interfacial charge transfer kinetics and/or surface capacitance of the working electrode 110.
[0088] The portable electrochemical reader 104 interfaced with the platform 102 applies the potential or current profiles to the working electrode 110 and records the resulting electrochemical responses. Further, the signal processing algorithms analyze the responses, compensating for any background signals or interferents, and convert them into quantitative measurements of LH levels.
[0089] The method further includes integrating or fabricating the biosensing device 100 as wearable or handheld personal care devices for real-time monitoring of luteinizing hormone. The integration process involves designing compact, ergonomic housings that accommodate the biosensing device 100 while ensuring user comfort and ease of use. For wearable applications, the biosensing apparatus 100 may be incorporated into devices such as smart watches, fitness bands, or adhesive patches, requiring miniaturization of the sensing platform and optimization of power consumption. The integration process also involves developing user-friendly interfaces and software applications that can interpret and display the biosensor data in real-time, providing easily understandable results and personalized insights. Connectivity features such as Bluetooth or Wi-Fi are implemented to enable seamless data transfer to mobile devices or cloud platforms for long-term tracking and analysis. The integrated devices are configured to be compatible with various sample collection methods, such as micro-fluidic channels for urine samples or minimally invasive techniques for interstitial fluid sampling, depending on the specific form factor. Additionally, the integration process includes incorporating data encryption and privacy measures to ensure the security of sensitive health information. Rigorous testing is conducted to ensure the integrated devices maintain the high performance standards of the biosensor while withstanding the demands of daily use. This seamless integration of the biosensing apparatus 100 into everyday devices marks a significant advancement in accessible and continuous hormone monitoring, empowering users with real-time health insights and facilitating proactive management of reproductive health.
[0090] Accordingly, the device and method of the present disclosure offers multiple advantages over conventional approaches by providing a highly sensitive, rapid, and user-friendly solution for luteinizing hormone detection. The use of the Pd-Ti₃C₂ MXene nanocomposite significantly enhances the electrochemical performance, resulting in improved sensitivity and a wide linear detection range from 1 to 200 mIU/mL. The low limit of detection (3 mIU/mL) and quantification (8 mIU/mL) enable accurate measurement of LH even at basal levels. The rapid detection time of 30 seconds represents a substantial improvement over traditional laboratory methods, facilitating real-time monitoring and immediate decision-making. The biosensor's long-term stability, maintaining performance for at least 1.5 months with minimal degradation, addresses a common challenge in biosensor technology and reduces the need for frequent recalibration or replacement. Its ability to function consistently across varying pH and storage conditions enhances its reliability in diverse testing environments. The flexibility and adaptability of the biosensing apparatus for integration into wearable and handheld devices open new possibilities for continuous, non-invasive hormone monitoring, potentially revolutionizing fertility tracking and management of hormonal disorders. Furthermore, the cost-effectiveness and scalability of the screen-printed electrode platform make this technology accessible for widespread use, including in resource-limited settings. By combining high analytical performance with practical usability, this biosensing approach bridges the gap between laboratory-grade accuracy and point-of-care convenience, offering a powerful tool for personalized women's health management and advancing the field of mobile health diagnostics.
[0091] The embodiments of the present invention disclosed herein are intended to be illustrative and not limiting. Other embodiments are possible and modifications may be made to the embodiments without departing from the spirit and scope of the invention. As such, these embodiments are only illustrative of the inventive concepts contained herein.
, Claims:1. A biosensing device (100) for sensitive and specific detection of luteinizing hormone (LH) (308), comprising:
an electrochemical transduction platform (102) comprising at least one electrode including a working electrode (110), a reference electrode (106), and a counter electrode (108);
a conductive nanostructured interface (304) deposited on the working electrode (110), the conductive nanostructured interface (304) comprising a titanium carbide (Ti₃C₂Tx) MXene nanocomposite functionalized with metallic nanoparticles; and
a biorecognition layer (306) comprising monoclonal anti-luteinizing hormone (anti-LH) antibodies immobilized onto the conductive nanostructured interface (304) via at least one immobilization mechanism selected from covalent coupling, non-covalent adsorption, cross-linking, or affinity-based interaction,
wherein the biosensing device (100) is configured to detect and quantify luteinizing hormone (308) upon exposure to a sample comprising luteinizing hormone (308), wherein the luteinizing hormone (308) specifically binds to immobilized anti-LH antibodies on the working electrode (110), thereby modulating interfacial charge transfer kinetics and/or surface capacitance of the working electrode (110), and such alterations being transduced into a measurable electrochemical signal, wherein magnitude of the electrochemical signal is quantitatively proportional to concentration of luteinizing hormone (308) in the sample.
2. The biosensing device (100) as claimed in claim 1, wherein the metallic nanoparticles are palladium nanoparticles, wherein the Pd nanoparticles are uniformly deposited onto a surface of Ti₃C₂ MXene, forming a palladium functionalized titanium carbide (Pd–Ti₃C₂) MXene nanocomposite, wherein the palladium nanoparticles are present in a form of nanodots, nanoclusters, or nanospheres.
3. The biosensing device (100) as claimed in claim 1, wherein the working electrode (110), the reference electrode (106), and the counter electrode (108) are configured in a screen-printed, flexible, or miniaturized format.
4. The biosensing device (100) as claimed in claim 1, wherein the working electrode (110) is fabricated from carbon-based conductive materials selected from graphite, graphene, carbon nanotubes, and carbon black.
5. The biosensing device (100) as claimed in claim 1, further comprising a portable electrochemical reader (104) operatively interfaced with the electrochemical transduction platform (102), the portable electrochemical reader (104) being configured to:
apply a controlled electrochemical potential and/or current to the working electrode (110);
acquire real-time electrochemical signals generated from biochemical interactions occurring at the surface of the working electrode (110);
digitize acquired analog signals via an onboard analog-to-digital converter (ADC);
process digitized signals through embedded firmware and/or software algorithms for baseline correction, noise filtering, and calibration; and
determine quantitative levels of luteinizing hormone (308) in the sample by correlating processed electrochemical signals with stored calibration models
wherein the portable electrochemical reader (104) optionally comprises a microcontroller, a rechargeable power source, a display unit, and/or a wireless communication module for enabling real-time, user-friendly, point-of-care detection of luteinizing hormone.
6. The biosensing device (100) as claimed in claim 1, wherein the biosensing device (100) is characterized by a linear detection range of 1–200 milli-international units per milliliter (mIU/mL) of luteinizing hormone and exhibits a limit of detection (LoD) of about 0.03 mIU/mL or lower.
7. The biosensing device (100) as claimed in claim 1, wherein the biosensing apparatus (100) is configured to be integrated into or fabricated as wearable or handheld personal care devices selected from wristbands, smart patches, smart watches, handheld diagnostic readers, or smartphone-integrated accessories.
8. The biosensing device (100) as claimed in claim 1, wherein the sample is any one of non-invasive sample or an invasive sample.
9. The biosensing device (100) as claimed in claim 1, wherein the biosensing device (100) exhibits high specificity toward luteinizing hormone (LH) with negligible cross-reactivity to potential interfering analytes including thyroid stimulating hormone (TSH), immunoglobulin G (IgG), uric acid (UA), and ascorbic acid (AA).
10. The biosensing device (100) as claimed in claim 1, wherein the biosensing device (100) demonstrates reproducibility with a relative standard deviation (RSD) of less than 5% and maintains long-term stability with a shelf life exceeding 50 days, exhibiting no more than about 3.2% degradation in electrochemical current response under optimized conditions.
11. The biosensing device (100) as claimed in claim 1, wherein the biosensing apparatus is configured to detect and quantify the luteinizing hormone within 30 seconds of sample introduction, as evidenced by rapid electrochemical signal generation in cyclic voltammetry profiles across a dynamic concentration range (1–200 mIU/mL).
12. A method for producing a biosensing device (100) for sensitive and specific detection of luteinizing hormone (LH) (308), comprising:
providing an electrochemical platform (102) comprising at least one electrode including a working electrode (110), a reference electrode (106), and a counter electrode (108);
depositing a conductive nanostructured interface (304) comprising a titanium carbide (Ti₃C₂Tx) MXene nanocomposite functionalized with metallic nanoparticles; and
immobilizing a biorecognition layer (306) comprising monoclonal anti-luteinizing hormone (anti-LH) antibodies onto conductive nanostructured interface (304) via at least one immobilization mechanism selected from covalent coupling, non-covalent adsorption, cross-linking, or affinity-based interaction.
13. The method as claimed in claim 12, wherein the metallic nanoparticles are palladium nanoparticles, wherein the Pd nanoparticles are uniformly deposited onto a surface of Ti₃C₂ MXene, forming a palladium functionalized titanium carbide (Pd–Ti₃C₂) MXene nanocomposite.
14. The method as claimed in claim 13, further comprising depositing the Pd nanoparticles uniformly onto a surface of Ti₃C₂ MXene, forming a palladium functionalized titanium carbide (Pd–Ti₃C₂) MXene nanocomposite on the surface of the working electrode (110), the deposition being carried out by drop-casting of pre-synthesized nanoparticles, wherein the palladium nanoparticles are present in a form of nanodots, nanoclusters, or nanospheres.
15. The method as claimed in claim 12, further comprising interfacing a portable electrochemical reader (104) with the electrochemical transduction platform (102), the portable electrochemical reader (104) being configured to:
apply a controlled electrochemical potential and/or current to the working electrode (110);
acquire real-time electrochemical signals generated from biochemical interactions occurring at the surface of the working electrode (110);
digitize acquired analog signals via an onboard analog-to-digital converter (ADC);
process digitized signals through embedded firmware and/or software algorithms for baseline correction, noise filtering, and calibration; and
determine quantitative levels of luteinizing hormone (308) in the sample by correlating processed electrochemical signals with stored calibration models,
wherein the portable electrochemical reader (104) optionally comprises a microcontroller, a rechargeable power source, a display unit, and/or a wireless communication module for enabling real-time, user-friendly, point-of-care detection of luteinizing hormone.
16. The method as claimed in claim 12, further comprising integrating or fabricating the biosensing device (100) as wearable or handheld personal care devices selected from wristbands, smart patches, smart watches, handheld diagnostic readers, or smartphone-integrated accessories.