WO2023057823A1

WO2023057823A1 - Detecting an antigen of a virus using electrochemical analysis

Info

Publication number: WO2023057823A1
Application number: PCT/IB2022/054574
Authority: WO
Inventors: Elias ALIPOUR; Hedayatollah Ghourchian; Mona SOROUSH
Original assignee: Alipour Elias; Hedayatollah Ghourchian; Soroush Mona
Priority date: 2021-10-10
Filing date: 2022-05-17
Publication date: 2023-04-13

Abstract

Disclosed herein is a system for detecting an antigen of a virus in a sample. The system includes an electrochemical cell, a magnetic-field generating device, a stimulator-analyzer device, and a processing unit. The electrochemical cell includes an electrically insulated container, an electrolyte solution, a working electrode, a counter electrode, and a reference electrode. The magnetic-field generating device includes an electrically conductive bar placed above the top surface of the working electrode. The stimulator-analyzer device is electrically connected to the electrodes. The processing unit includes a memory and a processor. The processor configures to perform a method of applying a set of voltages and measuring a produced set of currents using the stimulator-analyzer device, measuring a concentration of the antigen of the virus in the sample by measuring a maximum electrical current using an equation, and diagnosing a presence of the antigen in the sample.

Description

DETECTING AN ANTIGEN OF A VIRUS USING EEECTROCHEMICAE

ANAEYSIS

TECHNICAE FIELD

[0001] The present disclosure generally relates to a system and a method for detecting and/or quantifying an antigen of a virus in a sample, and more particularly, relates to a system and a method for detecting and/or quantifying hepatitis B antigen in a sample using electrochemical analysis.

BACKGROUND ART

[0002] Biosensor is an analytical device that is used for detection and quantification of a biological analyte in a sample by generating signals. Biosensors based on their mechanism of detection are classified as electrochemical, optical, thermal, and piezoelectric biosensors. Electrochemical biosensors can transduce biological events to electrical signals. Mechanism of electrochemical biosensors is based on measuring changes in conductance, resistance, or capacitance of electrochemical biosensor.

[0003] Immunosensors are a category of biosensors that may be used for detecting antigens and antibodies by immobilizing a bio-receptor (either an antigen or antibody) on physical transducers. Immunosensors use nanomaterials as a label for enhancing a signal produced by physical transducers. Immunosensors are vastly used in clinical diagnosis and monitoring of diseases. Immunosensors based on electrochemical recognition mechanisms may use three electrodes in an electrolyte media to analyze biological events.

[0004] Hepatitis B is an infectious disease caused by a virus called hepatitis B virus (HBV). The hepatitis virus can infect the liver and may cause death. HBV may cause chronic hepatitis and, eventually, liver failure, cirrhosis, and hepatocellular carcinoma. Therefore, diagnosis of HBV in early stages of the disease is one of the most critical global health problems. Diagnosing HBV in early stages may help to treat and reduce damages caused by HBV.

[0005] Immunosensors are vastly used to detect hepatitis B surface antigen (HBsAg) as a biomarker of HBV. Some known methods used by immunosensors for detecting HBsAg include qualitative or quantitative luminescence, amperometry, voltammetry and enzyme linked immunosorbent assay (ELISA) based methods. Ichiro Tono et al. presented a patent on “MEASURING SYSTEM USING OPTICALWAVEGUIDE, MEASURING DEVICE, MEASURING METHOD, OPTICALWAVEGUIDE TYPE SENSORCHIP, AND MAGNETIC FINE PARTICLE” (US 9.274,104 B2). Ichiro Tono et al. used light intensity difference between a surface containing antibody and a reference surface for analyzing samples. In another report, Thomas E. Rohr et al. presented a patent on “AGNETICALLY ASSISTED BINDING ASSAYS UTILIZING A MAGNETICALLY RESPONSIVE REAGENT” (5998224). Thomas E. Rohr et al. analyzed forces of samples containing antigen and reference samples to calculate a concentration of the antigen. These methods require markers/labels such as ELISA, horseradish peroxidase, etc., to diagnose an antigen of a virus in a sample.

[0006] There is, therefore, a need for a cost-effective, label-free, user-friendly, and fast method and system to quantitatively analyze HBV in a sample. There is further a need for a method and a system to detect and quantify HBVs in a sample with high accuracy.

SUMMARY OF THE DISCLOSURE

[0007] This summary is intended to provide an overview of the subject matter of this patent, and is not intended to identify essential elements or key elements of the subject matter, nor is it intended to be used to determine the scope of the claimed implementations. The proper scope of this patent may be ascertained from the claims set forth below in view of the detailed description below and the drawings.

[0008] According to one or more exemplary embodiments, the present disclosure is directed to a system for detecting an antigen of a virus in a sample. In an exemplary embodiment, an exemplary system may include an electrochemical cell, a magnetic field generating device, a stimulator-analyzer device, and a processing unit. In an exemplary embodiment, an exemplary electrochemical cell may include an electrically insulated container, an electrolyte solution poured inside an exemplary electrically insulated container, three electrodes being in contact with an exemplary electrolyte solution, and a suspension poured inside an exemplary electrolyte solution. In an exemplary embodiment, an exemplary three electrodes may include a working electrode, a counter electrode, and a reference electrode. In an exemplary embodiment, an exemplary working electrode may include a mercury substrate may be placed at an internal bottom surface of an exemplary electrically insulated container. In an exemplary embodiment, a first plurality of fragment antigen-binding (Fab) regions of antibodies of an exemplary virus may be deposited on a portion of top surface of an exemplary mercury substrate. In an exemplary embodiment, each respective Fab region of an antibody of an exemplary virus of an exemplary first plurality of Fab regions of exemplary antibodies of an exemplary virus may be configured to bind to a first part of an antigen of an exemplary virus in an exemplary sample. In an exemplary embodiment, an exemplary isolating material may cover parts of an exemplary top surface of an exemplary mercury substrate among an exemplary deposited first plurality of Fab regions of exemplary antibodies of an exemplary virus. In an exemplary embodiment, an exemplary counter electrode may be placed within an exemplary electrolyte solution. In an exemplary embodiment, an exemplary reference electrode may be placed within an exemplary electrolyte solution. In an exemplary embodiment, an exemplary suspension may be poured into an exemplary electrically insulated container. In an exemplary embodiment, an exemplary suspension may include a plurality of magnetic particles bound to a second plurality of Fab regions of exemplary antibodies of an exemplary virus. In an exemplary embodiment, each respective Fab region of an exemplary antibody of an exemplary virus of an exemplary second plurality of Fab regions of exemplary antibodies of an exemplary virus may be configured to bind to a second part of an exemplary antigen of an exemplary virus in an exemplary sample. In an exemplary embodiment, an exemplary magnetic field generating device may include an electrically conductive bar may be placed vertically along a longitudinal axis of an exemplary electrically conductive bar above an exemplary top surface of an exemplary working electrode. In an exemplary embodiment, a first end of an exemplary electrically conductive bar may be dipped into an exemplary electrolyte solution. In an exemplary embodiment, an exemplary magnetic field generating device may be configured to detach a complex from an exemplary top surface of an exemplary working electrode by applying a magnetic field to an exemplary electrolyte solution. In an exemplary embodiment, an exemplary complex may include a magnetic particle, a Fab region of an exemplary antibody of an exemplary virus of an exemplary second plurality of Fab regions of exemplary antibodies of an exemplary virus, an exemplary antigen of an exemplary virus, and a Fab region of an exemplary antibody of an exemplary virus of an exemplary first plurality of Fab regions of exemplary antibodies of an exemplary virus bound together. In an exemplary embodiment, an exemplary stimulator-analyzer device may be electrically connected to exemplary three electrodes. In an exemplary embodiment, an exemplary stimulator- analyzer device may be configured to apply a set of voltages between an exemplary reference electrode and an exemplary working electrode and measure a produced set of electrical currents between an exemplary working electrode and an exemplary counter electrode responsive to an exemplary applied set of voltages. In an exemplary embodiment, an exemplary processing unit may be electrically connected to an exemplary stimulator- analyzer device and an exemplary magnetic field generating device. In an exemplary embodiment, an exemplary processing unit may include a memory may have processor-readable instructions stored therein and a processor may be configured to access an exemplary memory and may execute exemplary processor-readable instructions, which, when executed by an exemplary processor may configure an exemplary processor to perform a method. In an exemplary embodiment, an exemplary method may include applying a set of voltages in a range of 50 mV to 220 mV between an exemplary reference electrode and an exemplary working electrode using an exemplary stimulatoranalyzer device, measuring a produced set of electrical currents between an exemplary counter electrode and an exemplary working electrode using an exemplary stimulator-analyzer device, measuring a maximum electrical current of an exemplary produced set of electrical currents, calculating a concentration of an exemplary antigen of an exemplary virus in an exemplary sample corresponding to an exemplary measured maximum electrical current of an exemplary produced set of electrical currents using an equation, an exemplary equation may include a mathematical relation between a plurality of maximum electrical currents and a respective plurality of antigen concentrations, and diagnosing a positive sample by detecting a presence of an exemplary antigen of an exemplary virus in an exemplary sample responsive to an exemplary measured concentration of an exemplary antigen of an exemplary virus may be more than a threshold concentration. In an exemplary embodiment, an exemplary antigen of an exemplary virus may include an antigen of hepatitis B virus with an exemplary threshold concentration of 1.2 ng/mL.

[0009] In an exemplary embodiment, an exemplary magnetic field generating device may further include an electrically conductive winding may be wrapped around an exemplary electrically conductive bar, and an electrical charge generator may be electrically connected to an exemplary electrically conductive bar and an exemplary electrically conductive winding from a second end of an exemplary electrically conductive bar. In an exemplary embodiment, an exemplary electrical charge generator may be configured to apply an electrical current in a range of 1 pA to 1 mA to an exemplary electrically conductive winding and an exemplary electrically conductive bar.

[0010] In an exemplary embodiment, an exemplary electrically conductive winding may be made of copper and an exemplary electrically conductive bar may be made of iron. In an exemplary embodiment, an exemplary first end of an exemplary electrically conductive bar may include a tip of a conical part of an exemplary electrically conductive bar. In an exemplary embodiment, an exemplary tip may be dipped inside an exemplary electrolyte solution. In an exemplary embodiment, a distance between an exemplary tip of an exemplary conical part and an exemplary top surface of an exemplary working electrode may be 1.25 mm or less.

[0011] In an exemplary embodiment, an exemplary system may further include a mercury source may be connected to an exemplary electrically insulated container via an electrically insulated tube. In an exemplary embodiment, a first end of an exemplary electrically insulated tube may be connected to a bottom surface of an exemplary electrically insulated container and a second end of an exemplary electrically insulated tube may be connected to an exemplary mercury source. In an exemplary embodiment, an exemplary mercury source and an exemplary electrically insulated tube may be configured to retain an amount of an exemplary mercury inside an exemplary electrically insulated container at a constant volume by charging an exemplary mercury from an exemplary mercury source to an exemplary electrically insulated container via an exemplary electrically insulated tube.

[0012] In an exemplary embodiment, an exemplary electrolyte solution may include at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof with a pH in a range of 6 to 9.

[0013] In an exemplary embodiment, an exemplary reference electrode may include an Ag/Ag Cl electrode. In an exemplary embodiment, an exemplary counter electrode may be made of platinum. In an exemplary embodiment, an exemplary isolating material may include 11- Mercaptoundecanoic acid (MUA).

[0014] In an exemplary embodiment, an exemplary suspension may include an exemplary plurality of magnetic particles bound to an exemplary second plurality of Fab regions of exemplary antibodies of an exemplary virus with a concentration of at least 25 pg/mL in at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof.

[0015] According to one or more exemplary embodiments, the present disclosure is directed to a method for detecting an antigen of a virus in a sample. In an exemplary embodiment, an exemplary method may include forming an electrochemical cell by forming a working electrode, adding an exemplary sample into an electrically insulated container in contact with an exemplary working electrode, pouring an electrolyte solution into an exemplary electrically insulated container, placing a counter electrode in contact with an exemplary electrolyte solution, placing a reference electrode in contact with an exemplary electrolyte solution, and electrically connecting an exemplary working electrode, an exemplary counter electrode, and an exemplary reference electrode to a stimulator-analyzer device. In an exemplary embodiment, forming an exemplary working electrode may include placing a mercury substrate at an internal bottom surface of an exemplary electrically insulated container, depositing a first plurality of fragment antigen-binding (Fab) regions of antibodies of an exemplary virus on a portion of top surface of an exemplary mercury substrate, and depositing an isolating material on an exemplary top surface of an exemplary mercury substrate. In an exemplary embodiment, each respective Fab region of an exemplary antibody of an exemplary virus of an exemplary first plurality of Fab regions of exemplary antibodies of an exemplary virus may be configured to bind to a first part of an exemplary antigen of an exemplary virus in an exemplary sample. In an exemplary embodiment, an exemplary isolating material may cover parts of an exemplary top surface of an exemplary mercury substrate among an exemplary deposited first plurality of Fab regions of exemplary antibodies of an exemplary virus. In an exemplary embodiment, an exemplary method may further include pouring a suspension into an exemplary electrically insulated container. In an exemplary embodiment, an exemplary suspension may include a plurality of magnetic particles bound to a second plurality of Fab regions of exemplary antibodies of an exemplary virus. In an exemplary embodiment, each respective Fab region of an exemplary antibody of an exemplary virus of an exemplary second plurality of Fab regions of exemplary antibodies of an exemplary virus may be configured to bind to a second part of an exemplary antigen of an exemplary virus in an exemplary sample. In an exemplary embodiment, an exemplary method may further include exposing an exemplary electrochemical cell to a magnetic field generating device. In an exemplary embodiment, exposing an exemplary electrochemical cell to an exemplary magnetic field generating device may include wrapping an electrically conductive winding around an electrically conductive bar, placing an exemplary electrically conductive bar with an exemplary wrapped electrically conductive winding above an exemplary top surface of an exemplary working electrode, and connecting an exemplary electrically conductive bar to an electrical charge generator from a second end of an exemplary electrically conductive bar. In an exemplary embodiment, a first end of an exemplary electrically conductive bar may be dipped inside an exemplary electrolyte solution. In an exemplary embodiment, an exemplary method may further include detaching a complex from an exemplary top surface of an exemplary working electrode by applying a magnetic field to an exemplary electrolyte solution using an exemplary magnetic field generating device. In an exemplary embodiment, an exemplary complex may include a magnetic particle, a Fab region of an antibody of an exemplary virus of an exemplary second plurality of Fab regions of exemplary antibodies of an exemplary virus, an exemplary antigen of an exemplary virus, and a Fab region of an exemplary antibody of an exemplary virus of an exemplary first plurality of Fab regions of exemplary antibodies of an exemplary virus bound together. In an exemplary embodiment, an exemplary method may further include applying a set of voltages in a range of 50 mV to 220 mV between an exemplary reference electrode and an exemplary working electrode using an exemplary stimulatoranalyzer device, measuring a produced set of electrical currents between an exemplary counter electrode and an exemplary working electrode using an exemplary stimulator-analyzer device, measuring a maximum electrical current of an exemplary produced set of electrical currents, calculating a concentration of an exemplary antigen of an exemplary virus in an exemplary sample corresponding to an exemplary measured maximum electrical current of an exemplary produced set of electrical currents using an equation, and diagnosing a positive sample by detecting a presence of an exemplary antigen of an exemplary virus in an exemplary sample responsive to an exemplary measured concentration of an exemplary antigen of an exemplary virus being more than a threshold concentration. In an exemplary embodiment, an exemplary equation may include a mathematical relation between a plurality of maximum electrical currents and a respective plurality of antigen concentrations. In an exemplary embodiment, an exemplary threshold concentration of hepatitis B virus may be 1.2 ng/mL.

[0016] In an exemplary embodiment, an exemplary method may further include generating an exemplary equation by measuring a set of reference maximum electrical currents. In an exemplary embodiment, measuring a set of reference maximum electrical currents may include forming a suspension of each reference solution of a plurality of reference solutions by adding predetermined concentrations of an exemplary antigen of an exemplary virus in at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof, adding an exemplary suspension of each reference solution of an exemplary plurality of reference solutions into an exemplary electrically insulated container in contact with an exemplary working electrode, heating an exemplary suspension of each reference solution of an exemplary plurality of reference solutions with an exemplary first plurality of Fab regions of exemplary antibodies of an exemplary virus may be deposited on an exemplary mercury substrate at a temperature in a range of 35°C to 40°C, forming an electrical connection between an exemplary counter electrode, an exemplary reference electrode, and an exemplary working electrode by pouring an exemplary electrolyte solution within an exemplary electrically insulated container , applying a magnetic field above an exemplary top surface of an exemplary working electrode using an exemplary magnetic field generating device, applying a set of voltages in a range from 50 mV to 220 mV between an exemplary reference electrode and an exemplary working electrode within each respective reference solution of an exemplary plurality of reference solutions using an exemplary stimulatoranalyzer device, measuring a produced set of electrical currents between an exemplary counter electrode and an exemplary working electrode associated with an exemplary suspension of each reference solution of an exemplary plurality of reference solutions responsive to an exemplary applied set of voltages, and measuring a maximum electrical current of each respective produced set of electrical currents may be associated with each respective reference solution of an exemplary plurality of reference solutions. In an exemplary embodiment, generating an exemplary equation may further include generating an exemplary equation by mathematically fitting an exemplary set of reference maximum electrical currents versus an exemplary respective concentrations of an exemplary antigen of an exemplary virus in an exemplary plurality of reference solutions.

[0017] In an exemplary embodiment, depositing an exemplary first plurality of Fab regions of exemplary antibodies of an exemplary virus on an exemplary portion of an exemplary top surface of an exemplary mercury substrate may include adding an exemplary first plurality of Fab regions of exemplary antibodies of an exemplary virus into an exemplary electrically insulated container with a concentration of at least 0.1 pg/mL in at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof. [0018] In an exemplary embodiment, depositing an exemplary first plurality of Fab regions of exemplary antibodies of an exemplary virus on an exemplary portion of an exemplary top surface of an exemplary mercury substrate may include heating an exemplary first plurality of Fab regions of exemplary antibodies of an exemplary virus deposited on an exemplary top surface of an exemplary mercury substrate in an oven at a temperature in a range of 35°C to 40°C for less than 12 hours. [0019] In an exemplary embodiment, pouring an exemplary suspension inside an exemplary electrolyte solution may further include heating an exemplary mixture of an exemplary second plurality of Fab regions of exemplary antibodies of an exemplary virus and an exemplary plurality of magnetic particles in an oven at a temperature in a range of 35°C to 40°C for 60 minutes to 120 minutes.

BRIEF DESCRIPTION OF THE DRAWINGS

[0020] The drawing figures depict one or more implementations in accord with the present teachings, by way of example only, not by way of limitation. In the figures, like reference numerals refer to the same or similar elements.

[0021] FIG. 1A illustrates a system for detecting an antigen of a virus in a sample, consistent with one or more exemplary embodiments of the present disclosure;

[0022] FIG. IB illustrates a magnified schematic view of an exemplary working electrode of an exemplary system for detecting an antigen of a virus in a sample, consistent with one or more exemplary embodiments of the present disclosure;

[0023] FIG. 1C illustrates a schematic view of a complex of magnetic particles functionalized by fragment antigen-binding (Fab) region of an antibody of a virus, Fab region of an antibody of an exemplary virus, and an antigen of an exemplary virus, consistent with one or more exemplary embodiments of the present disclosure;

[0024] FIG. 2 illustrates a computer system in which an embodiment of the present disclosure, or portions thereof, may be implemented as computer-readable code, consistent with one or more exemplary embodiments of the present disclosure;

[0025] FIG. 3A illustrates a flowchart of a method for detecting an antigen of a virus in a sample, consistent with one or more exemplary embodiments of the present disclosure;

[0026] FIG. 3B illustrates a flowchart of a method for forming an exemplary working electrode, consistent with one or more exemplary embodiments of the present disclosure;

[0027] FIG. 3C illustrates a flowchart of a method to generate an equation between a measured maximum electrical current and antigen concentration, consistent with one or more exemplary embodiments of the present disclosure;

[0028] FIG. 4 illustrates curves of produced currents versus time for a sample with HBsAg and a control sample, consistent with one or more exemplary embodiments of the present disclosure; [0029] FIG. 5 illustrates an optical density image of exemplary samples for analyzing activation of Fab regions immobilized on mercury substrate, consistent with one or more exemplary embodiments of the present disclosure;

[0030] FIG. 6 illustrates an optical density image of exemplary sample for analyzing activation of Fab regions attached to exemplary magnetic nanoparticles and magnetic beads, consistent with one or more exemplary embodiments of the present disclosure;

[0031] FIG. 7 illustrates an optical density image of exemplary samples for analyzing optimum Fab regions concentrations on mercury electrode, consistent with one or more exemplary embodiments of the present disclosure;

[0032] FIG. 8 illustrates an optical density image of exemplary samples for analyzing optimum Fab regions concentrations on magnetic beads, consistent with one or more exemplary embodiments of the present disclosure;

[0033] FIG. 9 illustrates produced currents versus an exemplary distance between an exemplary electrically conductive bar of a magnetic field generating device and an exemplary working electrode, consistent with one or more exemplary embodiments of the present disclosure;

[0034] FIG. 10 illustrates produced currents versus incubation time of depositing Fab regions on mercury substrate, consistent with one or more exemplary embodiments of the present disclosure;

[0035] FIG. 11 illustrates produced current versus incubation time for interaction between Fab regions and hepatitis B antigen, consistent with one or more exemplary embodiments of the present disclosure;

[0036] FIG. 12 illustrates produced current versus incubation time for interaction between an exemplary second plurality of Fab regions and hepatitis B antigen, consistent with one or more exemplary embodiments of the present disclosure;

[0037] FIG. 13 illustrates produced currents versus different temperatures for analyzing interaction between Fab regions and antigen, consistent with one or more exemplary embodiments of the present disclosure;

[0038] FIG. 14 illustrates produced currents versus different temperatures for analyzing interaction between Fab regions immobilized on magnetic particles and antigen, consistent with one or more exemplary embodiments of the present disclosure; [0039] FIG. 15 illustrates an image of optical density versus different pHs for analyzing interaction between Fab regions and mercury, consistent with one or more exemplary embodiments of the present disclosure;

[0040] FIG. 16 illustrates an image of optical density versus different pHs for analyzing interaction between Fab regions and magnetic particles, consistent with one or more exemplary embodiments of the present disclosure;

[0041] FIG. 17 illustrates produced currents versus different pHs of an exemplary system, consistent with one or more exemplary embodiments of the present disclosure;

[0042] FIG. 18 illustrates produced currents versus different concentrations of exemplary electrolyte solutions, consistent with one or more exemplary embodiments of the present disclosure;

[0043] FIG. 19 illustrates a calibration curve based on produced current versus different concentrations of HBsAg, consistent with one or more exemplary embodiments of the present disclosure; and

[0044] FIG. 20 illustrates produced current versus different samples, consistent with one or more exemplary embodiments of the present disclosure.

DESCRIPTION OF EMBODIMENTS

[0045] In the following detailed description, numerous specific details are set forth by way of examples in order to provide a thorough understanding of the relevant teachings. However, it should be apparent that the present teachings may be practiced without such details. In other instances, well known methods, procedures, components, and/or circuitry have been described at a relatively high-level, without detail, in order to avoid unnecessarily obscuring aspects of the present teachings.

[0046] The novel features which are believed to be characteristic of the present disclosure, as to its structure, organization, use and method of operation, together with further objectives and advantages thereof, will be better understood from the following discussion. In the following detailed description, numerous specific details are set forth by way of examples in order to provide a thorough understanding of the relevant teachings. However, it should be apparent that the present teachings may be practiced without such details. In other instances, well known methods, procedures, components, and/or circuitry have been described at a relatively high- level, without detail, in order to avoid unnecessarily obscuring aspects of the present teachings. The following detailed description is presented to enable a person skilled in the art to make and use the methods and devices disclosed in exemplary embodiments of the present disclosure. For purposes of explanation, specific nomenclature is set forth to provide a thorough understanding of the present disclosure. However, it will be apparent to one skilled in the art that these specific details are not required to practice the disclosed exemplary embodiments. Descriptions of specific exemplary embodiments are provided only as representative examples. Various modifications to the exemplary implementations will be readily apparent to one skilled in the art, and the general principles defined herein may be applied to other implementations and applications without departing from the scope of the present disclosure. The present disclosure is not intended to be limited to the implementations shown, but is to be accorded the widest possible scope consistent with the principles and features disclosed herein.

[0047] The present disclosure is directed to exemplary embodiments of a system and a method to detect and/or quantify an antigen of a virus in a sample. In an exemplary embodiment, a presence of an antigen of a virus may be detected in a sample utilizing exemplary method and/or system disclosed herein. In an exemplary embodiment, a concentration or an amount of an antigen of a virus may be detected utilizing exemplary method and/or system. In an exemplary embodiment, an exemplary antigen of a virus may include an antigen of hepatitis B virus (HBV). In an exemplary embodiment, an exemplary sample may include a fluid acquired from a human, or an animal. In an exemplary embodiment, an exemplary sample may be blood plasma.

[0048] An exemplary system may include an electrochemical cell, a magnetic field generating device, a stimulator analyzer device, and a processing unit. In an exemplary embodiment, an exemplary electrochemical cell may include an electrically insulated container. In an exemplary embodiment, an electrolyte solution with a pH in a range of 6 to 9 may be poured into an exemplary electrically insulated container. In an exemplary embodiment, an exemplary electrolyte solution may include at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof. In an exemplary embodiment, an exemplary electrochemical cell may further include a working electrode, a reference electrode, and a counter electrode. In an exemplary embodiment, exemplary electrodes may be in contact with an exemplary electrolyte solution within an exemplary electrically insulated container. In an exemplary embodiment, an exemplary electrolyte solution may form an electrical connection between exemplary electrodes. In an exemplary implementation, an exemplary working electrode may include a mercury substrate placed in an internal bottom of an exemplary electrically insulated container. In an exemplary embodiment, an exemplary working electrode may further include a first plurality of fragment antigen-binding (Fab) regions of antibodies of a virus deposited on a portion of an exemplary top surface of an exemplary mercury substrate. In an exemplary embodiment, an exemplary virus may be hepatitis B virus. As used herein, Fab region may refer to a region of an antibody that may bind to an antigen. In an exemplary embodiment, each Fab region of an exemplary antibody of an exemplary virus among an exemplary first plurality of Fab regions of antibodies of an exemplary virus may bind to a first part of an exemplary antigen of an exemplary virus in an exemplary sample. In an exemplary embodiment, an isolating material may cover parts of an exemplary top surface of an exemplary mercury substrate remained bare among an exemplary deposited first plurality of Fab regions of antibodies of an exemplary virus. In an exemplary embodiment, an exemplary isolating material may include 11-Mercaptoundecanoic acid (MUA). In an exemplary embodiment, each molecule of MUA may be smaller than each Fab region of an exemplary antibody of an exemplary virus. In an exemplary embodiment, an exemplary isolating material may be an electrically insulating material. In an exemplary embodiment, an exemplary isolating material may form an electrically insulating layer over exemplary bare parts of top surface of an exemplary mercury substrate among an exemplary deposited first plurality of Fab regions of exemplary antibodies of an exemplary virus.

[0049] In an exemplary embodiment, after depositing an exemplary isolating layer on an exemplary top surface of an exemplary mercury substrate, an exemplary sample may be added into an exemplary electrically insulated container. In an exemplary embodiment, an exemplary sample may be suspected to have an exemplary antigen of an exemplary virus therein. In an exemplary embodiment, an exemplary sample may be acquired from a person suspected to be infected by an exemplary virus. In an exemplary embodiment, a first part of an exemplary antigen of an exemplary virus may be bound to an exemplary first plurality of Fab regions of exemplary antibodies of an exemplary virus.

[0050] In an exemplary embodiment, a suspension of a plurality of magnetic particles bound to a second plurality of Fab regions of exemplary antibodies of an exemplary virus in at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof may be added into an exemplary electrically insulated container. In an exemplary embodiment, each magnetic particle of an exemplary plurality of magnetic particles may have a size in a range of 30 nm to 1 jam. In an exemplary embodiment, each respective Fab region of an exemplary antibody of an exemplary virus of an exemplary second plurality of Fab regions of exemplary antibodies of an exemplary virus may be bound to a second part of an exemplary antigen of an exemplary virus in an exemplary sample. In an exemplary embodiment, an exemplary virus may include hepatitis B virus.

[0051] In an exemplary embodiment, an exemplary counter electrode may be a rod- shaped electrode. In an exemplary embodiment an exemplary reference electrode may be a rod-shaped electrode. In an exemplary embodiment, an exemplary counter electrode may be placed within an exemplary electrolyte solution in a way that a first end of an exemplary counter electrode may be out of an exemplary electrolyte solution allowing for forming electrical connections. In an exemplary embodiment, an exemplary reference electrode may be placed within an exemplary electrolyte solution in a way that a first end of an exemplary reference electrode may be out of an exemplary electrolyte solution allowing for forming electrical connections.

[0052] In an exemplary embodiment, an exemplary magnetic field generating device may include an electrically conductive winding, an electrically conductive bar, and an electrical charge generator. In an exemplary embodiment, an exemplary electrically conductive winding may be wrapped around an exemplary electrically conductive bar. In an exemplary implementation, an electrical current may be applied to an exemplary electrically conductive winding and an exemplary electrically conductive bar using an exemplary electrical charge generator. In an exemplary implementation, an exemplary electrical current may be applied to an exemplary electrically conductive winding and an exemplary electrically conductive bar in a range of 1 p A to 1 mA to produce magnetic field. In an exemplary embodiment, an exemplary electrical charge generator may be electrically connected to an exemplary electrically conductive winding and an exemplary electrically conductive bar. In an exemplary implementation, an exemplary electrically conductive bar may be placed vertically along a longitudinal axis of an exemplary electrically conductive bar above an exemplary top surface of an exemplary working electrode. In an exemplary embodiment, an exemplary electrically conductive bar may be made of iron. In an exemplary embodiment, an exemplary electrically conductive bar may include a first end and a second end. In an exemplary embodiment, an exemplary second end of an exemplary electrically conductive bar may be electrically connected to an exemplary electrical charge generator. In an exemplary embodiment, an exemplary first end of an exemplary conductive bar may be placed within an exemplary electrolyte solution. In an exemplary embodiment, an exemplary first end of an exemplary electrically conductive bar may include a tip of a conical part of the electrically conductive bar. In an exemplary embodiment, an exemplary tip may be dipped inside an exemplary electrolyte solution. In an exemplary implementation, an exemplary tip may include a flat end point with a diameter in a range of 1mm to 1.2 mm. In an exemplary embodiment, an exemplary flat end point of an exemplary tip may be placed in a distance of less than 1.25 mm to an exemplary top surface of an exemplary working electrode. In exemplary embodiment, an exemplary magnetic field generating device may be used for detaching a complex from an exemplary top surface of an exemplary mercury substrate by applying an exemplary magnetic field to an exemplary electrolyte solution. In an exemplary embodiment, an exemplary complex may include a magnetic particle, a Fab region of an exemplary antibody of an exemplary virus of an exemplary second plurality of Fab regions of exemplary antibodies of an exemplary virus, an exemplary antigen of an exemplary virus, and a Fab region of an exemplary antibody of an exemplary virus of an exemplary first plurality of Fab regions of exemplary antibodies of an exemplary virus bound together. In an exemplary embodiment, detaching an exemplary complex from an exemplary top surface of an exemplary mercury substrate may form a vacant space on an exemplary top surface of an exemplary mercury substrate. In an exemplary embodiment, exposed top surface of an exemplary mercury substrate may form an electrical connection with an exemplary counter electrode and an exemplary reference electrode. In an exemplary embodiment, after a while, an exemplary exposed top surface of an exemplary mercury substrate may be filled with an exemplary isolating material and an exemplary first plurality of Fab regions of exemplary antibodies of an exemplary virus. Filling an exemplary exposed top surface of an exemplary mercury substrate may happen due to fluidic nature of mercury.

[0053] In an exemplary embodiment, an exemplary stimulator-analyzer device may be electrically connected to an exemplary working electrode, an exemplary counter electrode, and an exemplary reference electrode. In an exemplary embodiment, an exemplary stimulatoranalyzer device may be used to apply a set of voltages between an exemplary reference electrode and an exemplary working electrode in a range of 50 mV to 220 mV. Afterward, a set of electrical currents produced between an exemplary counter electrode and an exemplary working electrode may be measured utilizing an exemplary stimulator analyzer device. In an exemplary embodiment, an exemplary set of voltages may be applied after applying an exemplary magnetic field to an exemplary electrolyte solution. In an exemplary embodiment, applying an exemplary magnetic field may detach an exemplary complex from an exemplary top surface of an exemplary mercury substrate. Therefore, an exemplary set of electrical currents may be produced after applying an exemplary set of voltages utilizing an exemplary stimulator-analyzer device. In an exemplary embodiment, an exemplary produced set of electrical currents may decrease when an exemplary bare top surface of an exemplary mercury substrate is filled.

[0054] In an exemplary embodiment, an exemplary processing unit may be electrically connected to an exemplary stimulator- analyzer device and an exemplary electrical charge generator of an exemplary magnetic field generating device. In an exemplary embodiment, an exemplary processing unit may be a computer system. In an exemplary embodiment, an exemplary processing unit may include a memory and a processor. In an exemplary embodiment, an exemplary memory may include processor-readable instructions stored therein. In an exemplary embodiment, an exemplary processor may access an exemplary memory and execute exemplary processor-readable instructions, which, when executed by an exemplary processor may configure an exemplary processor to perform a method. In an exemplary embodiment, an exemplary method may include applying a set of voltages in a range of 50 mV to 220 mV between an exemplary reference electrode and an exemplary working electrode and applying an exemplary magnetic field to an exemplary electrolyte solution. After applying an exemplary set of voltages utilizing an exemplary stimulator-analyzer device, a set of electrical currents may be measured between an exemplary working electrode and an exemplary counter electrode utilizing an exemplary stimulator-analyzer device. In an exemplary embodiment, an exemplary set of electrical currents may be measured after applying an exemplary magnetic field utilizing an exemplary magnetic field generating device by applying an electrical current to an exemplary electrically conductive bar and an exemplary electrically conductive winding utilizing an exemplary electrical charge generator. In an exemplary embodiment, a concentration of an exemplary antigen of an exemplary virus in an exemplary sample may be measured utilizing an exemplary processing unit. In an exemplary embodiment, an exemplary concentration of an exemplary antigen of an exemplary virus in an exemplary sample may be measured by measuring/detecting a maximum produced electrical current of an exemplary produced set of electrical currents of an exemplary antigen of an exemplary virus. In an exemplary embodiment, an exemplary concentration of an exemplary antigen of an exemplary virus in an exemplary sample may be calculated using an equation based on an exemplary measured maximum produced electrical current. In an exemplary embodiment, a positive sample may be diagnosed by detecting a presence of an exemplary antigen of an exemplary virus in an exemplary sample if an exemplary measured concentration of an exemplary antigen of an exemplary virus is more than a threshold concentration. In an exemplary embodiment, an exemplary threshold concentration of hepatitis B antigen may be 1.2 ng/mL of hepatitis B antigen in an exemplary sample.

[0055] FIG. 1A illustrates a system 100 for detecting an antigen of a virus in a sample, consistent with one or more exemplary embodiments of the present disclosure. In an exemplary embodiment, system 100 may be utilized by methods 300 and 317 illustrated herein below. In an exemplary embodiment, system 100 may be used to detect and quantify an antigen of a virus in a sample. In an exemplary embodiment, a concentration or amount of an exemplary antigen of an exemplary virus may be detected utilizing system 100. In an exemplary embodiment, system 100 may include an electrochemical cell, a magnetic field generating device, a processing unit 104, and a stimulator- analyzer device 102. In an exemplary embodiment, an exemplary electrochemical cell may include an electrically insulated container 130, a counter electrode 114, a reference electrode 116, a working electrode 134, and an electrolyte solution 126. In an exemplary embodiment, electrolyte solution 126 may be poured within electrically insulated container 130. In an exemplary embodiment, electrolyte solution 126 may form electrical connections between counter electrode 114, reference electrode 116, and working electrode 134. In an exemplary embodiment, electrolyte solution 126 may include at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof. In an exemplary embodiment, electrolyte solution 126 may have a pH in a range of 6 to 9. An exemplary pH range of 6 to 9 may have compatibility with pH of human body. In an exemplary embodiment, counter electrode 114 and reference electrode 116 may have a cylindrical configuration. In an exemplary embodiment, a first end 114a of counter electrode 114 may be electrically connected to stimulator-analyzer device 102 using electrical connection 112. In an exemplary embodiment, a second end 114b of counter electrode 114 may be dipped inside electrolyte solution 126. In an exemplary embodiment, a first end 116a of reference electrode 116 may be electrically connected to stimulator- analyzer device 102 using electrical connection 110. In an exemplary embodiment, a second end 116b of reference electrode 116 may be dipped inside electrolyte solution 126. FIG. IB illustrates a magnified schematic view of working electrode 134 of system 100 for detecting an antigen of a virus in a sample, consistent with one or more exemplary embodiments of the present disclosure. In an exemplary embodiment, working electrode 134 may include a mercury substrate 144, a first plurality of first fragment antigen-binding (Fab) regions 140 of exemplary antibodies of an exemplary virus, and an isolating material 142. In an exemplary embodiment, mercury substrate 144 may be supplied by a mercury source 136. In an exemplary embodiment, mercury source 136 may be connected to electrically insulated container 130 using an electrically insulated tube 132. In an exemplary embodiment, a first end 132a of electrically insulated tube 132 may be connected to a bottom surface of electrically insulated container 130. In an exemplary embodiment, a second end 132b of electrically insulated tube 132 may be connected to mercury source 136. In an exemplary embodiment, mercury source 136 and electrically insulated tube 132 may be configured to retain an amount of an exemplary mercury inside electrically insulated container 130 at a constant volume by charging an exemplary mercury from mercury source 136 to electrically insulated container 130 via electrically insulated tube 132. In an exemplary embodiment, electrically insulated tube 132 may be made of at least one of polymer, glass, and combinations thereof.

[0056] In an exemplary embodiment, first plurality of first Fab regions 140 may be prepared from an antibody of an exemplary virus. As used herein, a “Fab region” may refer to a region of an antibody that binds with antigens. In an exemplary embodiment, an exemplary antibody may include two parts. In an exemplary embodiment, exemplary two parts may include two sections of Fab region and one section of fragment crystallizable (Fc) region. In an exemplary embodiment, an enzyme may be used to separate Fab regions and Fc sections. In an exemplary embodiment, an exemplary enzyme may include pepsin. In an exemplary embodiment, two sections of Fab region may be separated by reducing exemplary two sections of Fab region.

[0057] In an exemplary embodiment, first Fab regions 140 of exemplary antibodies of an exemplary virus may cover parts of an exemplary top surface of mercury substrate 144. In an exemplary embodiment, first Fab regions 140 of an exemplary antibody of an exemplary virus may be used to bind selectively to a first part of an exemplary antigen of an exemplary virus. In an exemplary embodiment, an exemplary virus may include hepatitis B virus. In an exemplary embodiment, amine (NH2) functional groups on first Fab regions 140 may bind with mercury substrate 144. Therefore, mercury substrate 144 may be functionalized by first Fab regions 140 for interaction with an exemplary antigen of an exemplary virus. [0058] In an exemplary embodiment, isolating material 142 may be added to mercury substrate 144 functionalized by first Fab regions 140. In an exemplary embodiment, isolating material 142 may cover parts of an exemplary top surface of mercury substrate 144 among an exemplary deposited first plurality of first Fab regions 140 of an exemplary antibody of an exemplary virus. In an exemplary embodiment, isolating material 142 may include an electrically insulating material. In an exemplary embodiment, isolating material 142 may include 11-Mercaptoundecanoic acid (MUA). In an exemplary embodiment, isolating material 142 may form an electrically insulating layer between an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus. In an exemplary embodiment, due to smaller size of MUA in comparison with first Fab regions 140 of an exemplary antibody of an exemplary virus, a thin layer of MUA may tend to deposit among an exemplary first plurality of first Fab regions 140 of an exemplary antibody of an exemplary virus.

[0059] In an exemplary embodiment, an exemplary sample may be added into electrically insulated container 130 after depositing isolating material 142 and an exemplary first plurality of first Fab regions 140 of an exemplary antibody of an exemplary virus on an exemplary top surface of mercury substrate 144. In an exemplary embodiment, an exemplary sample may be suspected to have an exemplary antigen of an exemplary virus therein. In an exemplary embodiment, a first part of an exemplary antigen of an exemplary virus in an exemplary sample may bind to an exemplary first plurality of first Fab regions 140 of an exemplary antibody of an exemplary virus.

[0060] In an exemplary embodiment, working electrode 134 may further include a suspension added into electrically insulated container 130. In an exemplary embodiment, an exemplary suspension may include a plurality of magnetic particles. In an exemplary embodiment, exemplary particles may be bound to a second plurality of Fab regions of an exemplary antibody of an exemplary virus. FIG. 1C illustrates a schematic view of a complex of magnetic particles 148 functionalized by fragment antigen-binding (Fab) region 150 of an antibody of a virus, Fab regionl40 of an antibody of an exemplary virus, and an antigen of an exemplary virus, consistent with one or more exemplary embodiments of the present disclosure. In an exemplary embodiment, each respective second Fab regions 150 of an exemplary antibody of an exemplary virus of an exemplary second plurality of second Fab regions 150 of exemplary antibodies of an exemplary virus may be configured to bind to a second part of an exemplary antigen 146 of an exemplary virus in an exemplary sample. In an exemplary embodiment, magnetic particles 148 may have functionalized carboxyl groups. In an exemplary embodiment, exemplary carboxyl groups of magnetic particles 148 may react with amine groups of an exemplary second plurality of second Fab regions 150 of an exemplary antibody of an exemplary virus. In an exemplary embodiment, magnetic particles 148 may be fictionalized by an exemplary second plurality of second Fab regions 150 of an exemplary antibody of an exemplary virus. In an exemplary embodiment, an exemplary suspension may include an exemplary plurality of magnetic particles 148 bound to an exemplary second plurality of second Fab regions 150 of exemplary antibodies with a concentration of at least 25 pg/mL in at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof. In an exemplary embodiment, each magnetic particle 148 may include two second Fab regions 150 of an exemplary antibody of an exemplary virus for better interaction of magnetic particles 148 to antigen 146 of an exemplary virus. In an exemplary embodiment, each magnetic particle 148 of an exemplary plurality of magnetic particles may have a size in a range of 30 nm to 1 pm. In an exemplary embodiment, each magnetic particle 148 of an exemplary plurality of magnetic particles may have a spherical shape with a diameter in a range of 30 nm to 1 pm.

[0061] As illustrated in FIG. 1A, system 100 may further include an exemplary magnetic field generating device. In an exemplary embodiment, an exemplary magnetic field generating device may include an electrically conductive bar 118, an electrically conductive winding 122, and an electrical charge generator 106. In an exemplary embodiment, electrically conductive bar 118 may be placed vertically along a longitudinal axis 120 of electrically conductive bar 118 above an exemplary top surface of working electrode 134. In an exemplary embodiment, electrically conductive bar 118 may include two ends. In an exemplary embodiment, a first end 120a of electrically conductive bar 118 may be dipped within electrolyte solution 126. In an exemplary embodiment, a second end 120b of electrically conductive bar 118 may be electrically connected to electrical charge generator 106. In an exemplary embodiment, first end 120a of electrically conductive bar 118 may include a flat tip of a conical part 124 of electrically conductive bar 118. In an exemplary embodiment, conical part 124 may be a conical-shaped part of electrically conductive bar 118 having a flat base with a diameter in a range of 1 mm to 1.2 mm. In an exemplary embodiment, first end 120a may be dipped within electrolyte solution 126. In an exemplary embodiment, a portion of conical part 124 including first end 120a may be dipped within electrolyte solution 126. In an exemplary embodiment, conical part 124 may be entirely dipped within electrolyte solution 126. In an exemplary embodiment, a distance between first end 120a and working electrode 134 may be 1.25 mm or less.

[0062] In an exemplary embodiment, electrical charge generator 106 may be electrically connected to processing unit 104 using electrical connection 123. In an exemplary embodiment, electrical charge generator 106 may be configured to apply an electrical current to electrically conductive bar 118 and electrically conductive winding 122 in a range of 50 mV to 220 mV and generate a magnetic-field within electrolyte solution 126 responsive to the applied electrical current. In an exemplary embodiment, an exemplary magnetic field generated by an exemplary magnetic field generating device may be applied into electrolyte solution 126. In an exemplary embodiment, tip 124 of electrically conductive bar 118 may be dipped inside electrolyte solution 126. In an exemplary embodiment, an exemplary magnetic field generating device may be used for detaching a complex from an exemplary top surface of an exemplary mercury substrate. In an exemplary embodiment, an exemplary complex may include magnetic particle 148, second Fab regions 150 of an exemplary antibody of an exemplary virus of an exemplary second plurality of Fab regions of exemplary antibodies of an exemplary virus, antigen 146 of an exemplary virus, and first Fab regions 140 of an exemplary antibody of an exemplary virus of an exemplary first plurality of Fab regions of exemplary antibodies of an exemplary virus bound together.

[0063] As illustrated in FIG. 1A, system 100 may further include a stimulator-analyzer device 102 and a processing unit 104. In an exemplary embodiment, stimulator-analyzer device 102 may be electrically connected to first end 116a of reference electrode 116 utilizing electrical connection 110. In an exemplary embodiment, stimulator-analyzer device 102 may be electrically connected to first end 114a of counter electrode 114 utilizing electrical connection 112. In an exemplary embodiment, counter electrode 114 may include platinum electrode. In an exemplary embodiment, reference electrode 116 may include Ag/Ag Cl electrode. In an exemplary embodiment, stimulator-analyzer device 102 may apply a set of voltages in a range of 50 mV to 220 mV between reference electrode 116 and working electrode 134. In an exemplary embodiment, stimulator- analyzer device 102 may measure a produced set of electrical currents responsive to an exemplary applied set of voltages between working electrode 134 and counter electrode 114. [0064] In an exemplary embodiment, processing unit 104 may be electrically connected to stimulator-analyzer device 104 utilizing electrical connection 108. Furthermore, processing unit 104 may be electrically connected to electrical charge generator 106 utilizing electrical connection 123. In an exemplary embodiment, processing unit 104 may include a memory that may have processor-readable instructions stored therein. In an exemplary embodiment, processing unit 104 may further include a processor. In an exemplary embodiment, an exemplary processor may access an exemplary memory and may execute exemplary processor- readable instructions. In an exemplary embodiment, exemplary processor-readable instructions when executed by an exemplary processor may configure an exemplary processor to perform a method. In an exemplary embodiment, an exemplary method may include applying a set of voltage between reference electrode 116 and working electrode 134 using stimulator-analyzer device 102. In an exemplary embodiment, an exemplary set of voltages may be in a range of 50 mV to 220 mV. In an exemplary embodiment, an exemplary processor may measure a set of electrical currents between counter electrode 114 and working electrode 134 using stimulator-analyzer device 102. In an exemplary embodiment, an exemplary set of electrical currents may be measured after applying an exemplary magnetic field using an exemplary magnetic field generating device. In an exemplary embodiment, a concentration of antigen 146 of an exemplary virus in an exemplary sample may be measured using a maximum produced electrical current of an exemplary produced set of electrical currents via an equation. In an exemplary embodiment, an exemplary equation may be formed based on exemplary maximum electrical currents of an exemplary produced set of electrical currents and concentrations of suspensions of reference samples. In an exemplary embodiment, exemplary suspensions of exemplary reference samples may be prepared by adding predetermined concentrations of antigen 146 of an exemplary virus in at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof. In an exemplary embodiment, exemplary suspensions of exemplary reference samples may be analyzed using system 100. In an exemplary embodiment, maximum electrical currents produced between counter electrode 114 and working electrode 134 associated with exemplary suspensions of an exemplary plurality of reference samples may be plotted versus exemplary concentrations of exemplary suspensions of an exemplary plurality of reference samples. In an exemplary embodiment, an exemplary equation may be used to determine concentration of antigen 146 of an exemplary virus of unknown samples using processing unit 104. In an exemplary embodiment, processing unit 104 may be a computer system illustrated in FIG. 2 below.

[0065] In an exemplary embodiment, processing unit 104 may include a computer system. FIG. 2 illustrates a computer system 200 in which an embodiment of the present disclosure, or portions thereof, may be implemented as computer-readable code, consistent with one or more exemplary embodiments of the present disclosure. For example, steps 308 and 310 of flowchart 300 may be implemented in computer system 200 using hardware, software, firmware, tangible computer readable media having instructions stored thereon, or a combination thereof and may be implemented in one or more computer systems or other processing systems. Hardware, software, or any combination of such may embody any of the modules and components in FIG. 1A. In an exemplary embodiment, computer system 200 may include processor 204.

[0066] If programmable logic is used, such logic may execute on a commercially available processing platform or a special purpose device. One ordinary skill in the art may appreciate that an embodiment of the disclosed subject matter can be practiced with various computer system configurations, including multi-core multiprocessor systems, minicomputers, mainframe computers, computers linked or clustered with distributed functions, as well as pervasive or miniature computers that may be embedded into virtually any device.

[0067] For instance, a computing device having at least one processor device and a memory may be used to implement the above-described embodiments. A processor device may be a single processor, a plurality of processors, or combinations thereof. Processor devices may have one or more processor “cores.”

[0068] An embodiment of the invention is described in terms of this example computer system 200. After reading this description, it will become apparent to a person skilled in the relevant art how to implement the invention using other computer systems and/or computer architectures. Although operations may be described as a sequential process, some of the operations may in fact be performed in parallel, concurrently, and/or in a distributed environment, and with program code stored locally or remotely for access by single or multiprocessor machines. In addition, in some embodiments the order of operations may be rearranged without departing from the spirit of the disclosed subject matter.

[0069] Processor 204 may be a special purpose or a general-purpose processor device. As will be appreciated by persons skilled in the relevant art, processor 204 may also be a single processor in a multi-core/multiprocessor system, such system operating alone, or in a cluster of computing devices operating in a cluster or server farm. Processor 204 may be connected to a communication infrastructure 202, for example, a bus, message queue, network, or multicore message-passing scheme.

[0070] In an exemplary embodiment, computer system 200 may include a display interface 208, for example a video connector, to transfer data to a display unit 226, for example, a monitor. Computer system 200 may also include a main memory 206, for example, random access memory (RAM), and may also include a secondary memory 210. Secondary memory 210 may include, for example, a hard disk drive 212, and a removable storage drive 214. Removable storage drive 214 may include a floppy disk drive, a magnetic tape drive, an optical disk drive, a flash memory, or the like. Removable storage drive 214 may read from and/or write to a removable storage unit 224 in a well-known manner. Removable storage unit 224 may include a floppy disk, a magnetic tape, an optical disk, etc., which may be read by and written to by removable storage drive 214. As will be appreciated by persons skilled in the relevant art, removable storage unit 224 may include a computer usable storage medium having stored therein computer software and/or data.

[0071] In alternative implementations, secondary memory 210 may include other similar means for allowing computer programs or other instructions to be loaded into computer system 200. Such means may include, for example, a removable storage unit 222 and an interface 216. Examples of such means may include a program cartridge and cartridge interface (such as that found in video game devices), a removable memory chip (such as an EPROM, or PROM) and associated socket, and other removable storage units 222 and interfaces 216 which allow software and data to be transferred from removable storage unit 222 to computer system 200. [0072] Computer system 200 may also include a network interface 218. Network interface 218 allows software and data to be transferred between computer system 200 and external devices. Network interface 218 may include a modem, a network interface (such as an Ethernet card), a communications port, a PCMCIA slot and card, or the like. Software and data transferred via network interface 218 may be in the form of signals, which may be electronic, electromagnetic, optical, or other signals capable of being received by network interface 218. These signals may be provided to network interface 218 via a communications path 220. Communications path 220 carries signals and may be implemented using wire or cable, fiber optics, a phone line, a cellular phone link, an RF link or other communications channels. [0073] In this document, the terms “computer program medium” and “computer usable medium” are used to generally refer to media such as removable storage unit 224, removable storage unit 222, and a hard disk installed in hard disk drive 212. Computer program medium and computer usable medium may also refer to memories, such as main memory 306 and secondary memory 210, which may be memory semiconductors (e.g. DRAMs, etc.).

[0074] Computer programs (also called computer control logic) are stored in main memory 206 and/or secondary memory 210. Computer programs may also be received via network interface 218. Such computer programs, when executed, enable computer system 200 to implement different embodiments of the present disclosure as discussed herein. In particular, the computer programs, when executed, enable processor device 204 to implement the processes of the present disclosure, such as the operations in method 300 illustrated by flowchart 300 of FIG 3, discussed below. Accordingly, such computer programs represent controllers of computer system 200. Where an exemplary embodiment of method 300 is implemented using software, the software may be stored in a computer program product and loaded into computer system 200 using removable storage drive 214, interface 216, and hard disk drive 212, or network interface 218.

[0075] Embodiments of the present disclosure also may be directed to computer program products including software stored on any computer useable medium. Such software, when executed in one or more data processing device, causes a data processing device to operate as described herein. An embodiment of the present disclosure may employ any computer useable or readable medium. Examples of computer useable mediums include, but are not limited to, primary storage devices (e.g., any type of random access memory), secondary storage devices (e.g., hard drives, floppy disks, CD ROMS, ZIP disks, tapes, magnetic storage devices, and optical storage devices, MEMS, nanotechnological storage device, etc.).

[0076] The embodiments have been described above with the aid of functional building blocks illustrating the implementation of specified functions and relationships thereof. The boundaries of these functional building blocks have been arbitrarily defined herein for the convenience of the description. Alternate boundaries can be defined so long as the specified functions and relationships thereof are appropriately performed. In an exemplary implementation, exemplary system 100 may be configured to detect a virus antigen in a sample via measuring maximum produced electrical currents using an equation. Exemplary system 100 may be utilized by a method 300 for detection and/or quantification of antigens of a virus in a sample described herein below. In an exemplary embodiment, processing unit 104 may include a computer system similar to computer system 200.

[0077] According to one or more exemplary embodiments, the present disclosure is further directed to exemplary embodiments of a method for detecting and/or quantifying an antigen of a virus in a sample. In an exemplary embodiment, a concentration or amount of an antigen of a virus may be detected utilizing an exemplary method. FIG. 3A illustrates a flowchart of a method 300 for detecting an antigen of a virus in a sample, consistent with one or more exemplary embodiments of the present disclosure. In an exemplary embodiment, method 300 may include a step 302 of forming an electrochemical cell by placing three electrodes inside an electrically insulated container in contact with an electrolyte solution, the three electrodes comprising a working electrode, a counter electrode, and a reference electrode, a step 304 of pouring a suspension inside the electrically insulated container, a step 306 of exposing the electrochemical cell to a magnetic field generating device, a step 308 of measuring a maximum electrical current of a produced set of electrical currents between the working electrode and the counter electrode responsive to an applied set of voltages between the reference electrode and the working electrode using a stimulator-analyzer device, and a step 310 of diagnosing a positive sample by measuring a concentration of the antigen of the virus in the sample based on the measured maximum electrical current.

[0078] In an exemplary embodiment, step 302 of forming an exemplary electrochemical cell may include placing three electrodes in electrolyte solution 126. In an exemplary embodiment, an exemplary electrochemical cell may include electrically insulating container 130, three electrodes, and electrolyte solution 126. In an exemplary embodiment, electrolyte solution 126 may be poured inside an electrically insulated container 130. In an exemplary embodiment, electrolyte solution 126 may be an ionic liquid. In an exemplary embodiment, electrolyte solution 126 may include at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof. In an exemplary embodiment, three electrodes may include reference electrode 116, counter electrode 114, and working electrode 134. In an exemplary embodiment, second end 116b of reference electrode 116 may be dipped inside electrolyte solution 126. In an exemplary embodiment, second end 114b of counter electrode 114 may be dipped inside electrolyte solution 126. In an exemplary embodiment, reference electrode 116 may include a cylindrical shape. In an exemplary embodiment, reference electrode 116 may include Ag/AgCl electrode. In an exemplary embodiment, counter electrode 114 may include a cylindrical shape. In an exemplary embodiment, counter electrode 114 may be made of platinum. In an exemplary embodiment, working electrode 134 may include mercury substrate 144 may be placed on an exemplary bottom of electrically insulated container 130. FIG. 3B illustrates a flowchart of a method 311 for forming working electrode 134, consistent with one or more exemplary embodiments of the present disclosure In an exemplary embodiment, method 311 may include a step 312 of placing a mercury substrate at an internal bottom surface of the electrically insulated container, a step 314 of depositing a plurality of fragment antigen-binding (Fab) regions of antibodies of a virus on a portion of the top surface of the mercury substrate, and a step 316 of depositing an isolating material on the top surface of the mercury substrate.

[0079] In an exemplary embodiment, step 312 of placing mercury substrate 144 at an internal bottom surface of electrically insulated container 130 may include forming mercury substrate 144 using mercury source 136 via electrically insulated tube 132. In an exemplary embodiment, electrically insulated tube 132 may be connected from first end 132a of electrically insulated tube 132 to an exemplary bottom of electrically insulated container 130. In an exemplary embodiment, electrically insulated tube 132 may be connected to mercury source 136 from second end 132b of electrically insulated tube 132. In an exemplary embodiment, mercury source 136 may be used to retain an amount of an exemplary mercury inside electrically insolated container 130 at a constant volume by charging an exemplary mercury from mercury source 136 to electrically insolated container 130 via electrically insulated tube 132. In an exemplary embodiment, an exemplary top surface of mercury substrate 144 may include an area in a range of 1 mm² to 2 mm².

[0080] In an exemplary embodiment, step 314 of depositing an exemplary first plurality of Fab regions of exemplary antibodies of an exemplary virus on a portion of an exemplary top surface of an exemplary mercury substrate may include pouring a predetermined concentration of an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus solution into electrically insulated container 130. In an exemplary embodiment, an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus may be dissolved in a buffer solution. In an exemplary embodiment, an exemplary buffer solution may include at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof. In an exemplary embodiment, an exemplary buffer solution may have a pH in a range of 6 to 9. In an exemplary embodiment, a concentration of an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus solution may be in a range of 0.1 pg/mL to 0.5 pg/mL. In an exemplary embodiment, depositing an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus on a portion of an exemplary top surface of an exemplary mercury substrate may include heating an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus solution and mercury substrate 144 at a temperature in a range of 35°C to 40°C. In an exemplary embodiment, an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus solution and mercury substrate 144 may be heated in an oven. In an exemplary embodiment, an exemplary oven may include an incubator. As used herein an incubator is a device to carry out processes that require regulated temperature, humidity, pressure, etc. In an exemplary embodiment, an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus solution and mercury substrate 144 may be heated for less than 12 hours. In an exemplary embodiment, amine functional groups on an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus may form chemical bonds with an exemplary top surface of mercury substrate 144. In an exemplary embodiment, an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus may be deposited on a portion of an exemplary top surface of mercury substrate 144. In an exemplary embodiment, heating an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus solution and mercury substrate 144 may initiate a reaction between mercury substrate 144 and amine functional groups on an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus. In an exemplary embodiment, an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus solution and mercury substrate 144 may be heated in an incubator. In an exemplary embodiment, a concentration of an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus solution incubated with mercury substrate 144 may be in a range of 0.1 pg/mL to 0.5 pg/mL in an exemplary buffer solution. In an exemplary embodiment, 10 pg to 50 pg of an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus may be deposited on mercury substrate 144. In an exemplary embodiment, an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus may be used for interacting with a first part of antigens 146 of an exemplary virus. In an exemplary embodiment, after heating an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus solution and mercury substrate 144, an exemplary buffer solution may be added into electrically insulated container 130 to dissolve unreacted an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus. In an exemplary embodiment, an exemplary buffet solution may be used as a washing solvent to remove unreacted an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus. In an exemplary embodiment, an exemplary buffer may be extracted from electrically insulated container 130. In an exemplary embodiment, an exemplary buffer may include at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof. In an exemplary embodiment, a concentration of an exemplary buffer solution may be more than 500 mM.

[0081] In an exemplary embodiment, step 316 of depositing an isolating material on an exemplary top surface of mercury substrate 144 may include adding a predetermined amount of an exemplary isolating material into electrically insulated container 130. In an exemplary embodiment, depositing an exemplary isolating material on an exemplary top surface of mercury substrate 144 may include heating an exemplary isolating material with mercury substrate 144 after deposition an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus on an exemplary top surface of mercury substrate 144. In an exemplary embodiment, an exemplary predetermined amount of an exemplary isolating material added into electrically insulated container 130 may be in a range of 10 pg : 40 pg to 50 pg : 80 pg (isolating material: first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus). In an exemplary embodiment, an exemplary isolating material may include 11-Mercaptoundecanoic acid (MUA). In an exemplary embodiment, an exemplary isolating material may form an isolating layer 142 among an exemplary deposited first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus on an exemplary top surface of mercury substrate 144. In an exemplary embodiment, depositing an exemplary isolating material among an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus may include heating an exemplary isolating material and mercury substrate 144 at a temperature in a range of 35°C to 40°C. In an exemplary embodiment, an exemplary isolating material deposited on an exemplary top surface of mercury substrate 144 may be heated in an oven. In an exemplary embodiment, an exemplary oven may include an incubator. In an exemplary embodiment, an exemplary isolating material and mercury substrate 144 may be heated for 60 minutes to 120 minutes. In an exemplary embodiment, an exemplary isolating material may form isolating layer 142. In an exemplary embodiment, molecules of MUA may be smaller than first Fab regions 140 of exemplary antibodies of an exemplary virus. Therefore, exemplary molecules of MUA may be deposited among an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus. In an exemplary embodiment, isolating layer 142 may isolate mercury substrate 144 from electrolyte solution 126. In an exemplary embodiment, electrolyte solution 126 may be an ionic liquid. In an exemplary embodiment, electrolyte solution 126 may form an electrical connection between counter electrode 114, reference electrode 116, and working electrode 134. In an exemplary embodiment, electrolyte solution 126 may include at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof. In an exemplary embodiment, after heating an exemplary isolating material and mercury substrate 144, an exemplary buffer solution may be added into electrically insulated container 130 to dissolve unreacted an exemplary isolating material. In an exemplary embodiment, an exemplary buffet solution may be used as a washing solvent to remove unreacted an exemplary isolating material. In an exemplary embodiment, an exemplary buffer may be extracted from electrically insulated container 130. In an exemplary embodiment, an exemplary buffer may include at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof. In an exemplary embodiment, a concentration of an exemplary buffer solution may be more than 500 mM.

[0082] In an exemplary embodiment, after depositing isolating layer 142, an exemplary sample may be poured into electrically insulated container 130. In an exemplary embodiment, an exemplary sample may include blood plasma. In an exemplary embodiment, an exemplary sample may be suspected to have antigen 146 therein. In an exemplary embodiment, 100 pL of an exemplary sample may be added to 0.1 pg to 0.5 pg of an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus deposited on an exemplary top surface of mercury substrate 144. In an exemplary embodiment, an exemplary sample and an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus may be heated at a temperature in a range of 35°C to 40°C for 60 minutes to 120 minutes. In an exemplary embodiment, heating an exemplary sample may enhance a rate of a reaction between antigen 146 and an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus. In an exemplary embodiment, after heating an exemplary sample and an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus, an exemplary buffer solution may be added into electrically insulated container 130 to remove unreacted antigen 146. In an exemplary embodiment, a concentration of an exemplary buffer solution may be more than 500 mM. In an exemplary embodiment, an exemplary buffer may include at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof.

[0083] In an exemplary embodiment, step 304 of pouring a suspension into the electrically insulated container 130 may include forming a suspension of magnetic particles 148 in an exemplary buffer solution. In an exemplary embodiment, a concentration of an exemplary buffer solution may be more than 500 mM. In an exemplary embodiment, an exemplary buffer may include at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof. In an exemplary embodiment, magnetic particles 148 may be bound to an exemplary second plurality of second Fab regions 150 of exemplary antibodies of an exemplary virus. In an exemplary embodiment, an exemplary suspension added into electrically insulated container 130 may be heated at a temperature in a range of 35°C to 40°C. In an exemplary embodiment, an exemplary suspension may be heated for 60 minutes to 120 minutes. In an exemplary embodiment, magnetic particles 148 may be attached to an exemplary second plurality of second Fab regions 150 of exemplary antibodies of an exemplary virus. In an exemplary embodiment, an exemplary second plurality of second Fab regions 150 of exemplary antibodies of an exemplary virus may include two attachment sites for better interaction with a second part of antigens 146 of an exemplary virus. In an exemplary embodiment, a size of each magnetic particle 148 of an exemplary plurality of magnetic particles 148 may be in a range of 30 nm to 1 pm. In an exemplary embodiment, a concentration of an exemplary suspension may be at least 25 pg/mL in an exemplary buffer solution. In an exemplary embodiment, an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus may bind to a first part of antigen 146 of an exemplary virus. In an exemplary embodiment, an exemplary second plurality of second Fab regions 150 of exemplary antibodies of an exemplary virus may bind to a second part of antigen 146 of an exemplary virus. Results indicated that the best concentration of magnetic particles bound to an exemplary second plurality of second Fab regions 150 of exemplary antibodies of an exemplary virus in an exemplary buffer solution may be above 25 pg/ml.

[0084] In an exemplary embodiment, step 306 of exposing an exemplary electrochemical cell to an exemplary magnetic field generating device may include placing a magnetic field generating device above an exemplary top surface of working electrode 134 that may be accomplished for applying magnetic field into electrolyte solution 126. In an exemplary embodiment, an exemplary magnetic field generating device may include electrically conductive bar 118, electrically conductive winding 122, and electrical charge generator 106. In an exemplary embodiment, electrically conductive winding 122 may be wrapped around electrically conductive bar 118. In an exemplary embodiment, electrically conductive bar 118 may include first end 120a with a conical tip 124. In an exemplary embodiment, electrically conductive winding 122 may be wrapped around electrically conductive bar 118 except conical tip 124. In an exemplary embodiment, electrically conductive bar 118 may be made of iron. In an exemplary embodiment, electrically conductive winding 122 may be made of copper. In an exemplary embodiment, electrical charge generator 106 may be electrically connected to electrically conductive winding 122 and electrically conductive bar 118 using an electrical connection 128. In an exemplary embodiment, electrical charge generator 106 may apply electric current in a range of 1 pA to 1 mA to electrically conductive winding 122 and electrically conductive bar 118 via an electrical connection 128. In an exemplary embodiment, an exemplary magnetic field generating device may apply an exemplary magnetic field within electrolyte solution 126. In an exemplary embodiment, applying an exemplary magnetic field within electrolyte solution 126 may detach a complex. In an exemplary embodiment, an exemplary complex may include magnetic particle 148, second Fab regions 150 of an exemplary antibody of an exemplary virus of an exemplary second plurality of second Fab regions 150 of exemplary antibodies of an exemplary virus, antigen 146 of an exemplary virus, and first Fab regions 140 of an exemplary antibody of an exemplary virus of an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus bound together. In an exemplary embodiment, detaching a plurality of exemplary complexes may leave a plurality of vacant spaces on an exemplary top surface of mercury substrate 144. In an exemplary embodiment, an exemplary plurality of vacant cavities may form an electrical connection between mercury substrate 144 and electrolyte solution 126.

[0085] In an exemplary embodiment, an exemplary applied magnetic field produced by an exemplary magnetic field generating device may be controlled by changing at least one of exemplary turns of electrically conductive winding 122, an exemplary applied electrical current, a length of electrically conductive winding 122, and combinations thereof. In an exemplary embodiment, an exemplary applied magnetic field may be enhanced by at least one of increasing exemplary number of turns of electrically conductive winding 122, reducing a thickness of an exemplary wire used for electrically conductive winding 122, reducing an exemplary diameter of electrically conductive bar 118, increasing applied electric current, increasing a length of electrically conductive winding 122, and combinations thereof.

[0086] In an exemplary embodiment, conical tip 124 may include a flat end point of first end 120a of electrically conductive bar 118 with a diameter in a range of 1mm to 1.2 mm. In an exemplary embodiment, electrically conductive bar 118 may be placed vertically along longitudinal axis 120 of electrically conductive bar 118 above an exemplary top surface of working electrode 134. In an exemplary embodiment, conical tip 124 of electrically conductive bar 118 may be dipped inside electrolyte solution 126. In an exemplary embodiment, a distance between conical tip 124 of electrically conductive bar 118 and an exemplary top surface of working electrode 134 may be less than 1.25 mm. In an exemplary embodiment, an exemplary flat end point (tip) of electrically conductive bar 118 may induce a homogenous field due to an exemplary structure and capability for being polished. In an exemplary embodiment, adjusting an exemplary distance between an exemplary flat end point of electrically conductive bar 118 and an exemplary top surface of working electrode 134 may be applicable when using an exemplary flat end point for electrically conductive bar 118. In an exemplary embodiment, adjusting an exemplary distance between an exemplary flat end point of electrically conductive bar 118 and an exemplary top surface of working electrode 134 may enhance controlling an exemplary intensity of an exemplary applied magnetic field. In an exemplary embodiment, an exemplary magnetic field may increase by decreasing an exemplary distance between an exemplary flat end point of electrically conductive bar 118 and an exemplary top surface of working electrode 134.

[0087] In an exemplary embodiment, at distances less than 1.25 mm between an exemplary flat end point of electrically conductive bar 118 and an exemplary top surface of working electrode 134, a rate of conductivity rise may enhance, but an amount of an exemplary enhancement of the electrical current may be constant. This is due to the similar concentration of antigen 146 in an exemplary sample. The reason may be that the same amount of magnetic particles 148 may be detached from working electrode 134 surface. In an exemplary embodiment, at different distances between an exemplary flat end point of electrically conductive bar 118 and an exemplary top surface of working electrode 134, the separation speed of magnetic particles 148 may be different. In an exemplary embodiment, at longer distances than 1.25 mm between an exemplary flat end point of electrically conductive bar 118 and an exemplary top surface of working electrode 134, an amount of magnetic field applied to magnetic particles 148 may be very low and may have no effect on magnetic particles 148. In an exemplary embodiment, when an exemplary distance between an exemplary flat end point of electrically conductive bar 118 and an exemplary top surface of working electrode 134 may reach 3 mm, an exemplary process of separation of an exemplary complex from an exemplary top surface of working electrode 134 may stop.

[0088] In an exemplary embodiment, step 308 of measuring a maximum electrical current of a produced set of electrical currents between working electrode 134 and counter electrode 114 responsive to applied set of voltages may include electrically connecting stimulator analyzer device 102 to exemplary electrodes. In an exemplary embodiment, stimulator analyzer device 102 may be electrically connected to working electrode 134 using electrical connection 135. In an exemplary embodiment, stimulator analyzer device 102 may be electrically connected to reference electrode 116 using electrical connection 110. In an exemplary embodiment, stimulator analyzer device 102 may be electrically connected to counter electrode 114 using electrical connection 112. In an exemplary embodiment, stimulator analyzer device 102 may apply a set of voltages in a range of 50 mV to 220 mV between working electrode 134 and reference electrode 116. In an exemplary embodiment, a set electrical currents may be produced responsive to an exemplary applied set of voltages. In an exemplary embodiment, an exemplary set of produced electrical currents may be measured between working electrode 134 and counter electrode 114. In an exemplary embodiment, an exemplary method of measuring produced set of electrical currents may be an amperometry method. As used herein, amperometry is a method that measures electrical currents versus time at a constant voltage. In an exemplary embodiment, an exemplary set of voltages may be measured for a time period in a range of 400 s to 600 s. In an exemplary embodiment, an exemplary produced set of electrical currents may be plotted versus time. In an exemplary embodiment, after applying an exemplary magnetic field within electrolyte solution 126, exemplary complexes may be detached from an exemplary top surface of mercury substrate 144. In an exemplary embodiment, applying an exemplary set of voltages between reference electrode 116 and working electrode 134 may produce an exemplary set of electrical current between working electrode 134 and counter electrode 114. In an exemplary embodiment, producing an exemplary set of electrical currents may show that antigen 146 may be presented in an exemplary sample. In an exemplary embodiment, a maximum intensity of an exemplary produced set of electrical currents may show quantity of antigen 146 in an exemplary sample. In an exemplary embodiment, when an exemplary quantity of antigen 146 in an exemplary sample increases, an exemplary maximum produced set of electrical currents may increase. In an exemplary embodiment, an exemplary produced set of electrical currents may be produced when exemplary complexes may be detached from an exemplary top surface of mercury substrate 144. In an exemplary embodiment, detaching exemplary complexes from an exemplary top surface of mercury substrate 144 may form vacant spaces on an exemplary top surface of mercury sunbathe 144. [0089] In an exemplary embodiment, after applying an exemplary magnetic field within electrolyte solution 126, Hg-Hg bonds may be broken due to weaker binding energy of Hg-Hg in comparison to Hg-Fab bonds. The bond-dissociation energy for C-C, C-N, antibody-antigen may be 618.3 kJ/mol, 750 kJ/mol, and 60.1 kJ/mol, respectively. The equivalent force for dissociation of C-C, C-N, antibody- antigen bonds may be 1600 Newton, 1940 Newton and 160xl0 ¹² Newton, respectively. The bond-dissociation energy for Hg-S and Hg-Hg are 217.3 kJ/mol and 8.10 kJ/mol or 562 Newton and 21xl0 ¹² Newton, respectively. Therefore, when a magnetic field may be applied, an exemplary complex may break from Hg-Hg bond (instead of breaking from the antigen-antibody interaction bond) and may separate from an exemplary top surface of mercury substrate 144. In an exemplary embodiment, this may create vacant cavities on an exemplary top surface of mercury substrate 144. In an exemplary embodiment, creating exemplary vacant cavities may cause significant changes in an exemplary conductivity and capacitance of system 100. By measuring changes in an exemplary conductance, a concentration of antigen 146 may be measured via an equation. In an exemplary embodiment, a minimum force of 10 ¹² Newton may be required to detach an exemplary complex from an exemplary top surface of mercury substrate 144.

[0090] In an exemplary embodiment, step 310 of diagnosing a positive sample by measuring a concentration of antigen 146 of an exemplary virus in an exemplary sample may include diagnosing a positive sample using processing unit 104. In an exemplary embodiment, processing unit 104 may be electrically connected to stimulator- analyzer device 102 using electrical connection 108. In an exemplary embodiment, processing unit 104 may be computer system 200 illustrated in FIG. 2. In an exemplary embodiment, processing unit 104 may include a memory and a processor. In an exemplary embodiment, an exemplary memory may have processor-readable instructions stored therein. In an exemplary embodiment, an exemplary processor may be configured to access an exemplary memory. In an exemplary embodiment, an exemplary processor may execute exemplary processor-readable instructions, which, when may be executed by an exemplary processor may configure an exemplary processor to perform a method. In an exemplary embodiment, an exemplary method may include applying a set of voltages in a range of 50 mV to 220 mV between reference electrode 116 and working electrode 134 using stimulator- analyzer device 102. In an exemplary embodiment, a produced set of electrical currents may be measured between counter electrode 114 and working electrode 134 using stimulator- analyzer device 102. In an exemplary embodiment, an exemplary produced set of electrical currents may be measures after applying an exemplary magnetic field within electrolyte solution 126 using an exemplary magnetic field generating device. In an exemplary embodiment, an equation may be formed based on exemplary maximum electrical currents of an exemplary produced set of electrical currents versus concentrations of antigen 146 in a plurality of reference samples. In an exemplary embodiment, an exemplary plurality of reference samples may be analyzed using system 100. FIG. 3C illustrates a flowchart of a method 317 to generate an equation between a measured maximum electrical current and antigen concentration, consistent with one or more exemplary embodiments of the present disclosure. In an exemplary embodiment, method 317 may include a step 318 of adding a suspension of each reference solution of a plurality of reference solutions into an electrically insulated container, a step 320 of heating the suspension of each reference solution of the plurality of reference solutions, a step 322 of applying a magnetic field above top surface of a working electrode, a step 324 of applying a set of voltages in a range from 50 mV to 220 mV between a reference electrode and the working electrode , a step 326 of measuring a produced set of electrical currents between a counter electrode and the working electrode, and a step 328 of generating an equation by mathematically fitting the equation based on maximum electrical currents of the produced set of electrical currents.

[0091] In an exemplary embodiment, step 318 of adding a suspension of each reference solution of a plurality of reference solutions into electrically insulated container 130 may include forming suspension of each reference solution of a plurality of reference solutions. In an exemplary embodiment, forming a suspension of each reference solution of a plurality of reference solutions may include adding a predetermined concentration of a plurality of predetermined concentrations of antigen 146 of an exemplary virus in an exemplary buffer solution. In an exemplary embodiment, an exemplary buffer may include at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof. In an exemplary embodiment, a concentration of an exemplary buffer solution may be more than 500 mM. In an exemplary embodiment, a suspension of each reference solution of a plurality of reference solutions may be added into electrically insulated container 130.

[0092] In an exemplary embodiment, step 320 of heating the suspension of each reference solution of the plurality of reference solutions may include heating an exemplary suspension of each reference solution of an exemplary plurality of reference solutions in an oven. In an exemplary embodiment, an exemplary suspension of each reference solution of an exemplary plurality of reference solutions may be heated at a temperature in a range of 35°C to 40°C.

[0093] In an exemplary embodiment, step 322 of applying a magnetic field may include applying an exemplary magnetic field above an exemplary top surface of working electrode 134 using an exemplary magnetic field generating device. In an exemplary embodiment, an exemplary magnetic field may be applied within electrolyte solution 126 containing predetermined concentrations of antigen 146. In an exemplary embodiment, applying an exemplary magnetic field may detach an exemplary complex of magnetic particle 148, second Fab regions 150 of an exemplary antibody of an exemplary virus of an exemplary second plurality of second Fab regions 150 of exemplary antibodies of an exemplary virus, antigen 146 of an exemplary virus, and first Fab regions 140 of an exemplary antibody of an exemplary virus of an exemplary first plurality of first Fab regions 140 of exemplary antibodies of an exemplary virus bound together from an exemplary top surface of mercury substrate 144.

[0094] In an exemplary embodiment, step 324 of applying a set of voltages may include applying an exemplary set of voltages in a range from 50 mV to 220 mV between reference electrode 116 and working electrode 134 using stimulator- analyzer device 102. In an exemplary embodiment, an exemplary set of voltages may be applied within each reference solution of an exemplary plurality of reference solutions.

[0095] In an exemplary embodiment, step 326 of measuring a produced set of electrical currents may include measuring an exemplary produced set of electrical currents between counter electrode 114 and working electrode 134. In an exemplary embodiment, an exemplary electrical current may be measured for 400 s to 600 s. In an exemplary embodiment, an exemplary electrical current of an exemplary plurality of reference samples may be measured using stimulator-analyzer device 102. In an exemplary embodiment, a maximum electrical current of each respective produced set of electrical currents associated with each respective reference solution of an exemplary plurality of reference solutions may be measured using processing unit 104.

[0096] In an exemplary embodiment, step 328 of generating an equation may include forming an exemplary equation based on exemplary maximum electrical currents associated with exemplary concentrations of an exemplary plurality of reference solutions. In an exemplary embodiment, exemplary maximum produced electrical currents associated with an exemplary plurality of reference samples may be plotted versus predetermined concentrations of an exemplary plurality of reference samples. In an exemplary embodiment, to determine a positive sample, a concentration of an unknown sample may be analyzed using system 100. In an exemplary embodiment, an exemplary concentration of an exemplary unknown sample may be measured using an exemplary equation via an exemplary maximum produced electrical current of an exemplary unknown sample. In an exemplary embodiment, an exemplary unknown sample may be declared positive when an exemplary concentration of antigen 146 of an exemplary unknown sample may be more than a threshold concentration of antigen 146. In an exemplary embodiment, when analyzing hepatitis B antigens in a sample, an exemplary threshold concentration of hepatitis B antigen may be 1.2 ng/mL.

[0097] Example 1: Detecting hepatitis B antigen in a sample

[0098] To detect hepatitis B antigens, a system similar to system 100 was used. For detecting hepatitis B antigen in a sample, a method similar to method 300 was used. To this end, 100 ng/mL of hepatitis B surface antigen (HBsAg) in phosphate buffer solution (PBS) was added to the sample and a control sample with no HBsAg was also used for comparison. FIG. 4 illustrates curves of produced currents versus time for a sample with HBsAg and a control sample, consistent with one or more exemplary embodiments of the present disclosure. After adding the sample containing HBsAg to the electrolyte solution, and applying the magnetic field, a complex of the HBsAg attached to magnetic particles and Fab regions deposited on the working electrode detached from the working electrode surface. Then a current between the working electrode, counter electrode, and the reference electrode forms which is proportional to a concentration of HBsAg. The beginning (406) of a peak in the curve (402) shows currents after applying the magnetic field. In the control sample with no HBsAg, after applying magnetic field, Fab regions deposited on the working electrode surface was still attached to the working electrode surface therefore no current change was noticed. Curve 402 shows produced currents versus time for the sample with HBsAg and curve 404 shows produced currents versus time for the control sample. In Example 1, the detachment of the complex may create some pinholes (vacant spaces) between the molecules of isolating layer. Therefore, a drastic change in the working electrode conductivity was noticed. After passing a few minutes, the conductivity reduces gradually which is due to filing pinholes with the mercury movement.

[0099] Results indicated that using an exemplary electrically conductive bar with radius of 0.5 cm, a length of 10 cm, a cone section length of 1 cm, and a cone section radius of 1 mm was an optimum dimensions for the electrically conductive bar. The distance between an exemplary flat end point of an exemplary electrically conductive bar and an exemplary top surface of an exemplary working electrode was 1 mm. An exemplary intensity of magnetic field was 0.25 tesla. In an exemplary embodiment, an intensity of an exemplary magnetic force applied on exemplary magnetic particles was 10 ¹² Newton. A magnitude of an exemplary applied electrical current to an exemplary magnetic field generating device was variable. A distance between an exemplary magnetic field generating device and an exemplary top surface of an exemplary working electrode was variable. An exemplary magnetic field applied on exemplary magnetic particles was enhanced by increasing an exemplary applied electrical current and decreasing an exemplary distance between an exemplary magnetic field generating device and an exemplary top surface of the working electrode.

[00100] An exemplary magnetic field generating device with 4000 turns of an exemplary electrically conductive winding and an exemplary electrically conductive bar with core diameter of 1 mm and an exemplary applied electrical current of 20 mA provided energy to detach an exemplary complex from the working electrode surface. The distance between the magnetic field generating device and the top surface of the working electrode was 1 mm. The electrically conductive bar was made of iron. The electrically conductive winding was made of copper. The produced magnetic fields were sufficient to detach exemplary complexes from the top surface of the working electrode.

[00101] Example 2: Estimation of the activity of Fab regions

[00102] In this Example a method for an accurate estimation of the activity of Fab regions embedded on mercury was used. Herein Fab region molecules are considered as the primary antibody. Fab region molecules were embedded on the mercury electrode, and then HBsAg was added into the working chamber. After an incubation time, the secondary antibody attached to the horseradish peroxidase (HRP) was added into the working chamber. After a while, a color change was observed. FIG. 5 illustrates an optical density image of exemplary samples for analyzing activation of Fab regions immobilized on mercury substrate, consistent with one or more exemplary embodiments of the present disclosure. Exemplary samples for analyzing activation of Fab regions include curve 502 for a sample containing mercury- immobilized Fab regions, HPR, HBsAg (positive sample control), curve 504 for a sample containing HPR and HBsAg, curve 506 for a sample with bare mercury electrode containing HPR and HBsAg, curve 508 for a sample containing HPR (negative sample), and curve 510 for a sample containing HRP.

[00103] Results indicated that Fab regions is active as the primary antibody on the mercury surface. The activity of antibody conjugated to magnetic beads was also investigated. Herein, antibody conjugated magnetic nanoparticles with 30 nm in diameter was considered as the primary antibody, then HBsAg was added into magnetic beads (MB) (particles with a size in a range of micrometers) and magnetic nanoparticles (MNP). After an incubation time the secondary antibody (AB) attached to the HRP was added into the magnetic nanoparticles, after a while, a color change was observed. FIG. 6 illustrates an optical density image of exemplary sample for analyzing activation of Fab regions attached to exemplary magnetic nanoparticles and magnetic beads, consistent with one or more exemplary embodiments of the present disclosure. Exemplary samples for analyzing activation of Fab regions attached to exemplary magnetic nanoparticles and magnetic beads include curve 602 for a sample containing magnetic nanoparticles and antibody, curve 604 for a sample containing magnetic beads and antibody, curve 606 for a sample containing magnetic particles (control), and curve 608 for a sample containing magnetic beads (control).

[00104] Example 3: Optimizing Fab regions concentration required for an exemplary

[00105] In Example 3, an experiment was performed to analyze optimum concentrations of Fab regions for preparing system 100. In Example 3, a method for optimizing the concentration of Fab regions for attachment to mercury electrode and magnetic beads using HPR was used. In an exemplary embodiment, the optimization method included incubation of Fab regions with concentrations of 0.1 pg/ml, 0.3 pg/ml, 0.5 pg/ml, 1 pg/ml, 5 pg/ml and 10 pg/ml separately on mercury electrode. FIG. 7 illustrates an optical density image of exemplary samples for analyzing optimum Fab regions concentrations on mercury electrode, consistent with one or more exemplary embodiments of the present disclosure. As shown in FIG. 7, the best concentrations for incubating Fab regions on the surface of mercury are concentrations above 0.5 pg/ml. To optimize the concentration of Fab regions antibodies for attachment to magnetic beads, the concentrations of 5 pg/ml, 10 pg/ml, 25 pg/ml, 50 pg/ml, 100 pg/ml and 200 pg/ml of secondary antibody conjugated magnetic particles were prepared in PBS and were tested independently. FIG. 8 illustrates an optical density image of exemplary samples for analyzing optimum Fab regions concentrations on magnetic beads, consistent with one or more exemplary embodiments of the present disclosure. Results indicated that the best concentration to form antibody conjugated magnetic bead is above 25 pg/ml.

[00106] Example 4: Optimizing an exemplary distance between an exemplary magnetic field generating device and an exemplary working electrode

[00107] In Example 3, an experiment was performed to optimize an exemplary distance between an exemplary magnetic field generating device and an exemplary working electrode for preparing system 100. Applying the magnetic field may lead to form holes in the isolating layer on the top surface of the mercury surface. Formation of holes may cause an increase in conductivity between the mercury electrode and the electrolyte solution. About 100 seconds after applying a magnetic field close to the mercury electrode, the magnetic particles may be detached from the surface of mercury electrode or capacitor. If this process is done correctly and quickly, exemplary holes may be created in the isolating layer that may increase the conductivity. FIG. 9 illustrates produced currents versus an exemplary distance between an exemplary electrically conductive bar of a magnetic field generating device and an exemplary working electrode, consistent with one or more exemplary embodiments of the present disclosure. As shown in FIG. 9, this process works well when the distance between the magnetic field generating device and the isolating layer on the mercury electrode is less than 1.25 mm. Shorter distances may be very effective. At distances less than 1.25 mm, an increase of conductance is faster, but the amount of the enhancement of the current is the same. This is due to the similar concentration of HBsAg in the sample and, therefore the same amount of magnetic particles that may be detached from the electrode surface. At different distances, the separation speed of these particles is different. At long distances (more than 1.25 mm), the amount of force-induced to magnetic beads is very low and may have no effect on the particles. When the distance is increased, the process of separation of the complex with magnetic particles is not completed eventually, and even at longer distances (about 3 mm), the process of separation of the complex almost stops. As shown in FIG. 9, the stronger the magnetic field is applied to the electrolyte solution by decreasing the distance between the magnetic field generating device and mercury electrode, the more significant the current change is observed before and after the separation of the magnetic particle complex.

[00108] Example 5: Optimizing incubation time for preparing an exemplary system for diagnosing hepatitis B

[00109] For optimizing incubation time for depositing Fab regions on mercury substrate, different time durations were tested. FIG. 10 illustrates produced currents versus incubation time of depositing Fab regions on mercury substrate, consistent with one or more exemplary embodiments of the present disclosure. To this end, 100 pL of a solution containing Fab regions may be poured on the surface of the mercury (working electrode). The mercury and the Fab regions were incubated for 1 hour, 2 hours, 3 hours, 6 hours, 12 hours, and 18 hours under the same condition. Then hepatitis B antigen was added into the modified electrode and then the secondary antibody conjugated with magnetic particles were added into the container. It was shown that the optimum time for immobilization of Fab regions on the mercury electrode surface was 12 hours. A shorter period of time can be applied for this stage because a high percentage of interaction between antigen and antibodies was occurred in the early moments of incubation. Gradually, the speed of the process of interaction between Fab regions and the mercury electrode was reached to a saturated stage.

[00110] For optimizing incubation time for interaction between hepatitis B antigen and Fab regions, another different time durations were tested. FIG. 11 illustrates produced current versus incubation time for interaction between Fab regions and hepatitis B antigen, consistent with one or more exemplary embodiments of the present disclosure. The Fab regions and hepatitis B antigen were incubated for 15 minutes, 30 minutes, 45 minutes, 60 minutes, and 120 minutes. The optimum incubation time for the interaction between Fab regions and hepatitis B antigen was 45 minutes.

[00111] For optimizing incubation time for interaction between hepatitis B antigen and second plurality of Fab regions immobilized on magnetic particles, another different time durations were tested. FIG. 12 illustrates produced current versus incubation time for interaction between an exemplary second plurality of Fab regions and hepatitis B antigen, consistent with one or more exemplary embodiments of the present disclosure. The second plurality of Fab regions and hepatitis B antigen were incubated for 15 minutes, 30 minutes, 45 minutes, 60 minutes, and 120 minutes. The optimum time for incubating the second plurality of Fab regions and hepatitis B antigen was 45 minutes. Another aspect of optimization of parameters affecting detection of HBsAg, was the effect of temperature on antibody-antigen interaction which was performed in two steps. One step was the interaction of Fab regions fragments of the first plurality of antibody with the antigen and the other was the interaction of the antigen with the second plurality of antibody conjugated to the magnetic particles.

[00112] Example 6: Optimizing incubation temperature for preparing an exemplary system for diagnosing hepatitis B

[00113] For optimizing incubation temperature for the interaction between Fab regions and antigen, different temperatures were tested. FIG. 13 illustrates produced currents versus different temperatures for analyzing interaction between Fab regions and antigen, consistent with one or more exemplary embodiments of the present disclosure. FIG. 14 illustrates produced currents versus different temperatures for analyzing interaction between Fab regions immobilized on magnetic particles and antigen, consistent with one or more exemplary embodiments of the present disclosure. The optimum temperature was obtained between 35°C and 40°C. The best temperature for interaction may be body temperature (37°C) which is expected to be the optimal temperature.

[00114] Example 7: Optimizing pH for preparing an exemplary system for diagnosing hepatitis B

[00115] For optimizing pH for the interaction between Fab regions on the top surface of mercury, different pHs were tested. The first plurality of Fab regions antibody was prepared in different PHs of 5, 6, 7, 8, 9 and 10 and was embedded into the mercury surface. And then, immobilized Fab regions antibody on mercury surface was incubated and washed. HRP was used as an indicator to analyze absorbance of samples in different pHs. To this end, a sample containing antigen, antibody, and HRP was used to be added into immobilized Fab regions on mercury at different pHs. FIG. 15 illustrates an image of optical density versus different pHs for analyzing interaction between Fab regions and mercury, consistent with one or more exemplary embodiments of the present disclosure. The optimum pH was achieved between 6 and 9.

[00116] For optimizing pH for the interaction between Fab regions and magnetic particles, different pHs were tested. Fab regions immobilized on magnetic particles was prepared in different PHs of 5, 6, 7, 8, 9 and 10. Phosphate buffer was used for controlling pH. FIG. 16 illustrates an image of optical density versus different pHs for analyzing interaction between Fab regions and magnetic particles, consistent with one or more exemplary embodiments of the present disclosure. The optimum pH was achieved between 8.5 and 9.5. FIG. 17 illustrates produced currents versus different pHs of an exemplary system, consistent with one or more exemplary embodiments of the present disclosure. Optimum pH for detection of HBsAg with an exemplary system is in a range between 6 and 9.

[00117] Example 8: Optimizing an exemplary electrolyte solution in an exemplary

[00118] For analyzing the performance of an exemplary system, three electrolyte solutions were used (PBS buffer, sodium chloride, and ferrocyanide buffer). Each of these buffers were used with different concentrations. FIG. 18 illustrates produced currents versus different concentrations of exemplary electrolyte solutions, consistent with one or more exemplary embodiments of the present disclosure. The results show that the optimal ion strength is obtained at concentrations above 500 mM and the best type of buffer to measure is ferrocyanide buffer.

[00119] Example 9: Optimizing the concentration of HBsAg

[00120] For optimizing the concentration of HBsAg, different concentrations of HBsAg were used to determine response of an exemplary system. FIG. 19 illustrates a calibration curve based on produced current versus different concentrations of HBsAg, consistent with one or more exemplary embodiments of the present disclosure. The current value (Al) indicates the difference between the response of an exemplary system before and after applying an external magnetic field. The change in Al is plotted against HbsAg concentration to determine the dynamic range of an exemplary system, and each point in FIG. 19 indicates the average value of three repetitions. Measurements were carried out in the presence of different concentrations of HBsAg after formation of exemplary complexes by using the second plurality of antibodies conjugated with magnetic beads. The responses are approximately linear for antigen concentrations from 0.1 to 40 ng.mL ¹, with the sensitivity (slope: s) of 3xl0^-7 A ng ¹ mL. However, the standard deviation at low concentration was ±9.1 x 10’⁹ A. Based on an exemplary system sensitivity and the standard deviations (SD), the detection limit (DL) of an exemplary system for detection of HBsAg was calculated to be 0.1 ng/mL.

[00121] Example 10: Analyzing selectivity of an exemplary system

[00122] To analyze selectivity of system 100, other samples including hepatitis C antigens (NS3 and NS4), bovine serum albumin (BSA), and hemoglobin were used. The concentration of all the samples were 100 ng/mL. FIG. 20 illustrates produced current versus different samples, consistent with one or more exemplary embodiments of the present disclosure. The experiments are carried out at room temperature. As seen in FIG. 20, a specific and repeatable response was observed for HBsAg, relative to the conductivity recorded for the other interfering samples.

[00123] System 100 was used for the detection of HBsAg concentration in real samples. Three measurements were performed for each blood sample without any pre-treatment. Corresponding concentration was calculated from the standard calibration curve obtained in Example 9. The concentration of HBsAg in the negative real samples were from 0.04 ng/mL to 0.1 ng/mL. The samples were also analyzed using HRP indicator for confirming the results. The concentration of HBsAg in positive samples were from 21 ng/mL to 40 ng/mL. The result for exemplary real samples are summarized in Table 1.

Table 1. Results of detecting HBsAg in exemplary real samples

Industrial Applicability

[00124] An exemplary system and method may be used for detecting and/or quantifying an antigen of a virus in a sample. An exemplary system and method may be used for accurate detection of hepatitis B antigen in blood samples. An exemplary method may be used in laboratories and hospitals due to fast and user friendly usage for detecting and/or quantifying an antigen of a virus such as hepatitis B in samples.

[00125] While the foregoing has described what are considered to be the best mode and/or other examples, it is understood that various modifications may be made therein and that the subject matter disclosed herein may be implemented in various forms and examples, and that the teachings may be applied in numerous applications, only some of which have been described herein. It is intended by the following claims to claim any and all applications, modifications and variations that fall within the true scope of the present teachings.

[00126] Unless otherwise stated, all measurements, values, ratings, positions, magnitudes, sizes, and other specifications that are set forth in this specification, including in the claims that follow, are approximate, not exact. They are intended to have a reasonable range that is consistent with the functions to which they relate and with what is customary in the art to which they pertain.

[00127] The scope of protection is limited solely by the claims that now follow. That scope is intended and should be interpreted to be as broad as is consistent with the ordinary meaning of the language that is used in the claims when interpreted in light of this specification and the prosecution history that follows and to encompass all structural and functional equivalents. Notwithstanding, none of the claims are intended to embrace subject matter that fails to satisfy the requirement of Sections 101, 102, or 103 of the Patent Act, nor should they be interpreted in such a way. Any unintended embracement of such subject matter is hereby disclaimed.

[00128] Except as stated immediately above, nothing that has been stated or illustrated is intended or should be interpreted to cause a dedication of any component, step, feature, object, benefit, advantage, or equivalent to the public, regardless of whether it is or is not recited in the claims.

[00129] It will be understood that the terms and expressions used herein have the ordinary meaning as is accorded to such terms and expressions with respect to their corresponding respective areas of inquiry and study except where specific meanings have otherwise been set forth herein. Relational terms such as first and second and the like may be used solely to distinguish one entity or action from another without necessarily requiring or implying any actual such relationship or order between such entities or actions. The terms “comprises,” “comprising,” or any other variation thereof, are intended to cover a nonexclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. An element proceeded by “a” or “an” does not, without further constraints, preclude the existence of additional identical elements in the process, method, article, or apparatus that comprises the element. [00130] The Abstract of the Disclosure is provided to allow the reader to quickly ascertain the nature of the technical disclosure. It is submitted with the understanding that it will not be used to interpret or limit the scope or meaning of the claims. In addition, in the foregoing Detailed Description, it can be seen that various features are grouped together in various implementations. This is for purposes of streamlining the disclosure, and is not to be interpreted as reflecting an intention that the claimed implementations require more features than are expressly recited in each claim. Rather, as the following claims reflect, inventive subject matter lies in less than all features of a single disclosed implementation. Thus, the following claims are hereby incorporated into the Detailed Description, with each claim standing on its own as a separately claimed subject matter.

[00131] While various implementations have been described, the description is intended to be exemplary, rather than limiting and it will be apparent to those of ordinary skill in the art that many more implementations and implementations are possible that are within the scope of the implementations. Although many possible combinations of features are shown in the accompanying figures and discussed in this detailed description, many other combinations of the disclosed features are possible. Any feature of any implementation may be used in combination with or substituted for any other feature or element in any other implementation unless specifically restricted. Therefore, it will be understood that any of the features shown and/or discussed in the present disclosure may be implemented together in any suitable combination. Accordingly, the implementations are not to be restricted except in light of the attached claims and their equivalents. Also, various modifications and changes may be made within the scope of the attached claims.

Claims

What is claimed is:

1. A system for detecting an antigen of a virus in a sample, comprising: an electrochemical cell, comprising: an electrically insulated container; an electrolyte solution poured inside the electrically insulated container; three electrodes being in contact with the electrolyte solution, the three electrodes comprising: a working electrode, comprising: a mercury substrate placed at an internal bottom surface of the electrically insulated container; a first plurality of fragment antigen-binding (Fab) regions of antibodies of the virus deposited on a portion of top surface of the mercury substrate, each respective Fab region of an antibody of the virus of the first plurality of Fab regions of the antibodies of the virus being configured to bind to a first part of the antigen of the virus in the sample; and an isolating material covering parts of the top surface of the mercury substrate among the deposited first plurality of Fab regions of the antibodies of the virus; a counter electrode placed within the electrolyte solution; and a reference electrode placed within the electrolyte solution; a suspension poured into the electrically insulated container, the suspension comprising a plurality of magnetic particles bound to a second plurality of Fab regions of the antibodies of the virus, each respective Fab region of an antibody of the virus of the second plurality of Fab regions of the

48 antibodies of the virus being configured to bind to a second part of the antigen of the virus in the sample; a magnetic field generating device comprising an electrically conductive bar placed vertically along a longitudinal axis of the electrically conductive bar above the top surface of the working electrode, a first end of the electrically conductive bar being dipped into the electrolyte solution, the magnetic field generating device configured to detach a complex from the top surface of the working electrode by applying a magnetic field to the electrolyte solution, the complex comprising a magnetic particle of the plurality of magnetic particles, a Fab region of the antibody of the virus of the second plurality of Fab regions of the antibodies of the virus, the antigen of the virus, and a Fab region of the antibody of the virus of the first plurality of Fab regions of the antibodies of the virus bound together, a stimulator-analyzer device electrically connected to the three electrodes, the stimulator- analyzer device being configured to: apply a set of voltages between the reference electrode and the working electrode; and measure a produced set of electrical currents between the working electrode and the counter electrode responsive to the applied set of voltages; and a processing unit electrically connected to the stimulator- analyzer device and the magnetic field generating device, the processing unit comprising: a memory having processor-readable instructions stored therein; and a processor configured to access the memory and execute the processor- readable instructions, which, when executed by the processor configures the processor to perform a method, the method comprising:

49 applying a set of voltages in a range of 50 mV to 220 mV between the reference electrode and the working electrode using the stimulatoranalyzer device; applying a magnetic field within the electrolyte solution using the magnetic field generating device; measuring a produced set of electrical currents between the counter electrode and the working electrode using the stimulator-analyzer device; measuring a maximum electrical current of the produced set of electrical currents; calculating a concentration of the antigen of the virus in the sample corresponding to the measured maximum electrical current of the produced set of electrical currents using an equation, the equation comprising a mathematical relation between a plurality of maximum electrical currents and a respective plurality of antigen concentrations; and diagnosing a positive sample by detecting a presence of the antigen of the virus in the sample responsive to the measured concentration of the antigen of the virus being more than a threshold concentration.

2. The system of claim 1, wherein the antigen of the virus comprises an antigen of hepatitis

B virus with the threshold concentration of 1.2 ng/mL.

3. The system of claim 1, wherein the magnetic field generating device further comprises:

50 an electrically conductive winding wrapped around the electrically conductive bar; and an electrical charge generator electrically connected to the electrically conductive bar and the electrically conductive winding from a second end of the electrically conductive bar, the electrical charge generator configured to apply an electrical current in a range of 1 pA to 1 mA to the electrically conductive winding and the electrically conductive bar.

4. The system of claim 3, wherein the electrically conductive winding is made of copper.

5. The system of claim 1, wherein the electrically conductive bar is made of iron.

6. The system of claim 1 , wherein the first end of the electrically conductive bar comprises a tip of a conical part of the electrically conductive bar, the tip being dipped inside the electrolyte solution, wherein a distance between the tip of the conical part and the top surface of the working electrode is 1.25 mm or less.

7. The system of claim 1, wherein the sample comprises blood plasma drawn from a person suspected to be infected by the antigen of the virus.

8. The system of claim 1, further comprising a mercury source connected to the electrically insulated container via an electrically insulated tube, a first end of the electrically insulated tube connecting to a bottom surface of the electrically insulated container and a

51 second end of the electrically insulated tube connecting to the mercury source, the mercury source and the electrically insulated tube being configured to: retain an amount of the mercury inside the electrically insulated container at a constant volume by charging the mercury from the mercury source to the electrically insulated container via the electrically insulated tube.

9. The system of claim 1, wherein a diameter of each magnetic particle of the plurality of magnetic particles is in a range of 30 nm to 1 pm.

10. The system of claim 1, wherein the electrolyte solution comprises at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof with a pH in a range of 6 to 9.

11. The system of claim 1, wherein the reference electrode comprises an Ag/Ag Cl electrode.

12. The system of claim 1, wherein the counter electrode is made of platinum.

13. The system of claim 1, wherein the isolating material comprises 11- Mercaptoundecanoic acid (MUA).

14. The system of claim 1, wherein the suspension comprises the plurality of magnetic particles bound to the second plurality of Fab regions of the antibodies of the virus with a concentration of at least 25 pg/mL in at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof. A method for detecting an antigen of a virus in a sample, the method comprising: forming an electrochemical cell, comprising: forming a working electrode, comprising: placing a mercury substrate at an internal bottom surface of an electrically insulated container; depositing a first plurality of fragment antigen-binding (Fab) regions of antibodies of the virus on a portion of top surface of the mercury substrate, each respective Fab region of an antibody of the virus of the first plurality of Fab regions of the antibodies of the virus being configured to bind to a first part of the antigen of the virus in the sample; and depositing an isolating material on the top surface of the mercury substrate, the isolating material covering parts of the top surface of the mercury substrate among the deposited first plurality of Fab regions of the antibodies of the virus; adding the sample into the electrically insulated container in contact with the working electrode; pouring an electrolyte solution into the electrically insulated container; placing a counter electrode in contact with the electrolyte solution; placing a reference electrode in contact with the electrolyte solution; electrically connecting the working electrode, the counter electrode, and the reference electrode to a stimulator-analyzer device; pouring a suspension into the electrically insulated container, the suspension comprising a plurality of magnetic particles bound to a second plurality of Fab regions of the antibodies of the virus, each respective Fab region of an antibody of the virus of the second plurality of Fab regions of the antibodies of the virus being configured to bind to a second part of the antigen of the virus in the sample; exposing the electrochemical cell to a magnetic field generating device, comprising: wrapping an electrically conductive winding around an electrically conductive bar; placing the electrically conductive bar with the wrapped electrically conductive winding above the top surface of the working electrode, a first end of the electrically conductive bar being dipped inside the electrolyte solution; and connecting the electrically conductive bar to an electrical charge generator from a second end of the electrically conductive bar; detaching a complex from the top surface of the working electrode by applying a magnetic field to the electrolyte solution using the magnetic field generating device, the complex comprising a magnetic particle, a Fab region of an antibody of the virus of the second plurality of Fab regions of the antibodies of the virus, the antigen of the virus, and a Fab region of the antibody of the virus of the first plurality of Fab regions of the antibodies of the virus bound together; applying a set of voltages in a range of 50 mV to 220 mV between the reference electrode and the working electrode using the stimulator- analyzer device; measuring a produced set of electrical currents between the counter electrode and the working electrode using the stimulator- analyzer device; measuring a maximum electrical current of the produced set of electrical currents;

54 calculating a concentration of the antigen of the virus in the sample corresponding to the measured maximum electrical current of the produced set of electrical currents using an equation, the equation comprising a mathematical relation between a plurality of maximum electrical currents and a respective plurality of antigen concentrations; and diagnosing a positive sample by detecting a presence of the antigen of the virus in the sample responsive to the measured concentration of the antigen of the virus being more than a threshold concentration.

16. The method of claim 15, wherein the method further comprises generating the equation, comprising: measuring a set of reference maximum electrical currents, comprising: forming a suspension of each reference solution of a plurality of reference solutions by adding a predetermined concentration of a plurality of predetermined concentrations of the antigen of the virus in at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof; adding the suspension of each reference solution of the plurality of reference solutions into the electrically insulated container in contact with the working electrode; heating the suspension of each reference solution of the plurality of reference solutions with the first plurality of Fab regions of the antibodies of the virus deposited on the mercury substrate at a temperature in a range of 35°C to

40°C;

55 forming an electrical connection between the counter electrode, the reference electrode, and the working electrode by pouring the electrolyte solution within the electrically insulated container; applying a magnetic field above the top surface of the working electrode using the magnetic field generating device; applying a set of voltages in a range from 50 mV to 220 mV between the reference electrode and the working electrode using the stimulator- analyzer device; measuring a produced set of electrical currents between the counter electrode and the working electrode associated with the suspension of each reference solution of the plurality of reference solutions responsive to the applied set of voltages; and measuring a maximum electrical current of each respective produced set of electrical currents associated with each respective reference solution of the plurality of reference solutions; and generating the equation by mathematically fitting the set of reference maximum electrical currents versus the respective concentrations of the antigen of the virus in the plurality of reference solutions.

17. The method of claim 15, wherein the antigen of the virus comprises an antigen of hepatitis B virus with the threshold concentration of 1.2 ng/mL.

18. The method of claim 15, wherein depositing the first plurality of Fab regions of the antibodies of the virus on the portion of the top surface of the mercury substrate comprises adding the first plurality of Fab regions of the antibodies of the virus into the electrically

56 insulated container with a concentration of at least 0.1 |ag/mL in at least one of a phosphate buffer solution, a sodium chloride solution, a ferrocyanide solution, and combinations thereof.

19. The method of claim 18, wherein depositing the first plurality of Fab regions of the antibodies of the virus on the portion of the top surface of the mercury substrate further comprises heating the first plurality of Fab regions of the antibodies of the virus deposited on the top surface of the mercury substrate in an oven at a temperature in a range of 35°C to 40°C for less than 12 hours.

20. The method of claim 15, wherein pouring the suspension into the electrically insulated container further comprises heating the mixture of the second plurality of Fab regions of the antibodies of the virus bound to the plurality of magnetic particles in an oven at a temperature in a range of 35°C to 40°C for 60 minutes tol20 minutes.

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