WO2025207778A1 - Sensor for rapid and sensitive quantification of sample characteristics - Google Patents

Abstract

An assay and device allow for rapid and effective detection of sample characteristics. The device of the present invention harnesses the detection power of an oximeter module. Light is passed through a sample and subsequently detected by the oximeter module, providing valuable information about the contents of the 5 sample. The device can be used for determining characteristics of a sample, including the presence of a target analyte. In one exemplary application, the device can be used for detection of target biomolecules, such as proteins, including, for example, vascular endothelial growth factor (VEGF), angiopoietin-2 (Ang-2), monocyte chemoattractant protein-1 (MCP-1), and interleukin-6 (IL-6).

Description

PATENT Page 1 P18228_02 SENSOR FOR RAPID AND SENSITIVE QUANTIFICATION OF SAMPLE CHARACTERISTICS CROSS REFERENCE TO RELATED APPLICATIONS [0001] This application claims the benefit of U.S. Provisional Application No.63/570,086, filed March 26, 2024, the content of each of the aforementioned applications is herein incorporated by reference in their entirety. FIELD OF THE INVENTION [0002] The present invention relates generally to detection devices. More particularly the present invention relates to a system for sensing of sample properties. BACKGROUND OF THE INVENTION [0003] About 30 million people in the US suffer from diabetic retinopathy (DR) or age- related macular degeneration (AMD), which can cause severe vision impairment or even blindness if not managed properly. These conditions are closely related to abnormal blood vessel growth in the retina, which is primarily driven by angiogenesis factors, particularly vascular endothelial growth factor (VEGF) or angiopoietin-2 (Ang-2); the levels of these factors are significantly higher in the aqueous humor of patients with DR or AMD compared to individuals with non-diabetic eye complications and are associated with the disease progression. Therefore, the first-line treatment is the intravitreal injection of angiogenesis inhibitors, such as antibodies targeting VEGF and/or Ang-2. Unfortunately, there are wide variations in therapeutic outcomes between patients, mainly due to inaccurate treatments. The pathogenesis of DR and AMD is mainly determined by angiogenesis factors, such as VEGF and Ang-2. Therefore, their levels in the aqueous humor can be a predictive biomarker for treatment response. Unfortunately, no method currently exists in a clinic to measure their PATENT Page 2 P18228_02 levels rapidly and sensitively, which has been a critical bottleneck for accurate patient stratification. [0004] Notably, inaccurate or delayed treatments result in permanent visual loss, and unnecessary injection of corticosteroids causes severe side effects, such as glaucoma. Moreover, the cost of intravitreal drug injection can be more than $20,000 per year for a patient. [0005] Therefore, there is an urgent clinical need for effective technologies to reveal which molecules need to be targeted for each patient before each treatment in a clinic. SUMMARY OF THE INVENTION In accordance with an embodiment of the present invention, a system is composed of an oximetry module wherein the oximetry module includes a light source, and wherein the oximetry module is configured to receive light from the light source after it is passed through a sample. The system also includes a microprocessor configured to receive data from the oximetry module. The microprocessor is also configured to determine characteristics about the sample. [0006] In accordance with another aspect of the present invention, the system includes a sample well. The system includes a shelf for receiving the sample well. The system includes a pair of oximetry modules in some embodiments, and in others the system includes four oximetry modules. The number of sample wells can be disposed in a sample tray, and the number of sample wells can correspond to the number of oximetry modules. The system is configured for detecting a target protein. The system is configured for detection of target proteins such as vascular endothelial growth factor (VEGF), angiopoietin-2 (Ang-2), PATENT Page 3 P18228_02 monocyte chemoattractant protein-1 (MCP-1), and interleukin-6 (IL-6). The system can be configured for detecting more than one target protein simultaneously. The microprocessor further comprises a display. The microprocessor is configured to transmit data wirelessly. The system includes a power source, and the power source can take the form of a battery. [0007] In accordance with another aspect of the present invention, a method for detecting characteristics of a sample includes applying light to the sample, wherein the light is a predetermined wavelength. The method includes detecting the light passed through the sample with an oximetry module. Further, the method includes processing data from the oximetry module with a microprocessor to determine characteristics of the sample. [0008] In accordance with another aspect of the present invention, the method includes displaying data about the sample on a display of the microprocessor. The method includes transmitting data from the microprocessor. The method includes wirelessly transmitting the data. The method includes detecting the presence of a target protein. The method includes detecting a concentration of a target protein. BRIEF DESCRIPTION OF THE FIGURES [0009] FIG.1A illustrates a perspective view of an immuno-magnetic, oximeter-based sensor, according to an embodiment of the present invention. [0010] FIG.1B illustrates a semi sectional view of an immuno-magnetic, oximeter-based sensor, according to an embodiment of the present invention. [0011] FIG.1C illustrates an exploded view of an immuno-magnetic, oximeter-based sensor, according to an embodiment of the present invention. PATENT Page 4 P18228_02 [0012] FIG.2 illustrates a schematic view of a protein sensing system and platform according to an embodiment of the present invention. [0013] FIGS.3A and 3B illustrate graphical views of performance between the present invention and conventional analytical technologies. [0014] FIG.4A-4D illustrate graphical views of detection of four different protein targets using iMOS. [0015] FIGS.5A and 5B illustrate graphical views of detection of spiked concentrations of target proteins in human aqueous humor. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS [0016] The presently disclosed subject matter now will be described more fully hereinafter with reference to the accompanying Drawings, in which some, but not all embodiments of the inventions are shown. Like numbers refer to like elements throughout. The presently disclosed subject matter may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will satisfy applicable legal requirements. Indeed, many modifications and other embodiments of the presently disclosed subject matter set forth herein will come to mind to one skilled in the art to which the presently disclosed subject matter pertains having the benefit of the teachings presented in the foregoing descriptions and the associated drawings. Therefore, it is to be understood that the presently disclosed subject matter is not to be limited to the specific embodiments disclosed and that modifications and other embodiments are intended to be included within the scope of the appended claims. [0017] An assay and device allow for rapid and effective analyte detection. The device can be used at point-of-care or in some instances an at-home test. The device of the present invention harnesses the detection power of an oximeter module. Light is passed through a PATENT Page 5 P18228_02 sample and subsequently detected by the oximeter module, providing valuable information, about the contents of the sample, such as the concentration of target analytes in a sample. The present invention can be used to detect target analytes in various applications known to or conceivable to one of skill in the art. Exemplary applications can include, but are not limited to detection of analytes in applications such as biomedicine, environments, and military. As long as target analytes are captured and reacted to generate an enzymatic signal, the sensors of the present invention can be applied. [0018] In one exemplary application, the device can be used for detection of vascular endothelial growth factor (VEGF), angiopoietin-2 (Ang-2), monocyte chemoattractant protein-1 (MCP-1), and interleukin-6 (IL-6). In this exemplary implementation, the device and method can be used to test aqueous humor samples from patients with DR or AMD before and after intravitreal injection of drugs that can detect VEGF and Ang-2 as low as 25 pg/mL within 90 min. While detection of VEGF and Ang-2 are described herein as an example, it is not to be considered limiting. Because, in some embodiments, the present invention measures the optical signals generated by enzymes attached to the target molecules, the device and method of the present invention can be applied more broadly to a variety of molecular detection, including proteins, metabolites, nucleic acids (e.g., DNAs or RNAs), toxins, heavy metals, and pathogens (e.g., virus and bacteria). [0019] An exemplary Immuno-Magnetic Oximeter-based Sensor (iMOS) of the present invention include 4 channels with a high sensitivity (<10 pg/mL) and a wide detection range (10-1000 pg/mL). Its detection power comes from an oximeter module, which is widely used in a portable fingertip pulse oximeter device. The present invention is the first to use the advantages of an oximetry module for biomolecular detection. PATENT Page 6 P18228_02 [0020] FIG.1A illustrates a perspective view of an immuno-magnetic, oximeter-based sensor, according to an embodiment of the present invention. FIG.1B illustrates a semi sectional view of an immuno-magnetic, oximeter-based sensor, according to an embodiment of the present invention. FIG.1C illustrates an exploded view of an immuno-magnetic, oximeter-based sensor, according to an embodiment of the present invention. The system 10 illustrated in FIG.1A shows a housing 12 for the components of the system 10. FIGS.1A and 1B also illustrate a microcontroller unit (MCU) 20 that includes a display 22. The display 22 can be used to display instruction and data related to the sample(s) being tested. [0021] As illustrated in FIG.1C, the system 10 includes an oximetry module printable circuit board (PCB) 14 that includes embedded oximetry units 16. As illustrated in FIG.1C, this exemplary system includes four embedded oximetry units 16. This is by way of example and the number of embedded oximetry units 16 included can be any number suitable to the testing being done. The embedded oximetry units 16 include LEDs. The LEDs in some embodiments have wavelengths of 650 nm and 800 nm. However, it should be noted that these wavelengths and the intensity of the light emitted from the LED can be adjusted and/or optimized for the type of testing and the desired results. The intensity of the LEDs can be changed and optimized so that a broader range of sample color intensities can be detected. Detection of a broader range of color intensities will allow detection of broader target analyte concentrations, in other words, a broader detection range. [0022] Further as illustrated in FIG.1C, a sample plate 24 is positioned below the oximetry module PCB 14. As illustrated in FIG.1C, the sample plate 24 includes four wells 26, which correspond to the number of embedded oximetry units 16. As noted above, the number of wells 26 can be increased or decreased based on the testing being done. While sample wells PATENT Page 7 P18228_02 are shown with respect to this exemplary embodiment, the sample can be disposed in any other container for inserting the sample into the system for detection known to or conceivable to one of skill in the art. [0023] The PCB of the exemplary embodiment shown in FIGS.1A-1C was developed to integrate four oximeter units with the (MCU). The developed PCB is the same size as the MCU, and each sample well in the loading plate is positioned right below each oximeter unit. This compact arrangement of the components, including a battery, allows for the development of a pocket-sized sensor. [0024] The MCU 20 is in communication with the PCB 14 and is configured to processes the data from the PCB 14 and its embedded oximetry modules 16, to detect and provide information at the point of care about the samples. In some exemplary embodiments, the system can include a mobile application for a smartphone/tablet for remote access/control of the system, such that the system can provide information about the biometric detection to users at the point of care. In such instances the MCU 20 can be configured for wireless or wired transmission of data. The system can also include a battery (not pictured) to provide power to the embedded oximetry modules 16 and the microprocessor 20. [0025] FIG.2 illustrates a schematic view of an analyte sensing system and platform according to an embodiment of the present invention. In some embodiments, the platform illustrated in FIG.2 can be referred to as an Immuno-Magnetic Oximeter-based Sensor (iMOS) platform. The iMOS platform includes an analyte quantification system 10 that integrates an assay with a custom-built oximeter-based absorbance sensor. In some embodiments, the assay can take the form of a conventional bead-based immunoassay. In an PATENT Page 8 P18228_02 exemplary embodiment for detecting biomolecules, the process begins with the collection of an aqueous humor sample (~50 μL), which is then mixed with immunoassay reagents containing magnetic beads coated with capture antibodies (cAb) and detector antibodies (dAb) conjugated with horseradish peroxidase (HRP) enzymes. Upon binding to the target biomolecule, these components form a ‘sandwich’ complex on the bead surface, with the quantity of formed complexes corresponding to the biomolecule concentration. Subsequently, a peroxidase chromogenic substrate, TMB (3,3',5,5'-tetramethylbenzidine), is added. The HRP enzyme oxidizes TMB, producing a color change that can be measured. The oxidized TMB absorbs light at approximately 650 nm, which is detected by the oximeter-based sensor in the biomolecule quantification system 10. The sensor transmits the absorbance data via Bluetooth^TM or other wireless data transmission protocol for real-time analysis. The iMOS platform is designed for rapid and quantitative diagnostics, capable of detecting various analytes. In some embodiments, the platform can be used as a point-of-care molecular diagnostic device for the detection of biomolecules, such as, but not limited to, protein biomarkers of vascular eye diseases; for example, the assay has already been established for four biomarkers: VEGF, Ang-2, IL-6, and MCP-1. These specific protein biomarkers are included by way of example and the system can be validated for the detection of any biomolecule or protein biomarkers known to one of skill in the art. With a total assay time of under 90 minutes, iMOS enables personalized diagnosis and treatment in a single visit to the clinic, which has not been possible before. [0026] FIGS.3A and 3B illustrate graphical views of performance between the present invention and conventional analytical technologies. FIG.3A illustrates that the ability to measure the absorbance of oxidized TMB was compared between the iMOS system and a conventional plate reader. A dilution series of HRP enzymes was reacted with TMB, and the PATENT Page 9 P18228_02 absorbance at 660 nm was measured. The readout curve of the iMOS system showed a shift to the left compared to that of a plate reader, indicating significant improvement in sensitivity. FIG.3B illustrates that different VEGF concentrations were analyzed using either the iMOS assay or a conventional enzyme-linked immunosorbent assay (ELISA). While both methods showed similar sensitivity, the iMOS assay required significantly less time than the ELISA (90 min vs.23 hours). [0027] FIG.4A-4D illustrate graphical views of detection of four different protein targets using iMOS. These four protein targets are included simply by way of examples of biomolecules that can be detected by the system of the present invention. It should be noted that any biomolecule known to or conceivable to one of skill in the art can be detected. Four different protein targets, VEGF, Ang-2, MCP-1, and IL-6 were successfully detected using iMOS with a limit of detection (LOD) of 1.8, 34.1, 16.0 and 21.6 pg/mL, respectively. The LOD was calculated using the following formula: _{[0028] 3} FIG.4A illustrates the detection of VEGF. FIG.4B illustrates the detection of Ang-2. FIG. 4C illustrates the detection of MCP-1, and FIG.4D illustrates the detection of IL-6. [0029] FIGS.5A and 5B illustrate graphical views of the detection of spiked concentrations of target biomolecules, and more specifically proteins in human aqueous humor. Different concentrations of VEGF recombinant proteins, as illustrated in FIG.5A and MCP-1 recombinant proteins, as illustrated in FIG.5B were spiked into human aqueous humor samples and quantified using iMOS. The measured concentrations closely matched the expected increases from the spiked amounts compared to the non-spiked control (Ctrl), demonstrating the system's accuracy. These results indicate that iMOS is well-suited for PATENT Page 10 P18228_02 detecting VEGF and MCP-1 in human aqueous humor, highlighting its compatibility with complex biological samples. [0030] In exemplary embodiments directed to eye health, the system of the present invention is designed to test 80-100 mL of aqueous humor from the eye to analyze the cytokines. The cytokine profile is provided within approximately 90 minutes. A doctor can then deliver more precise treatment to the patient. The sensitivity of the assay can be optimized with programming of the oximetry module as well as the microprocessor and user interface within the application. [0031] It should also be noted that herein the steps of the method described can be carried out using a microprocessor, a computer, a smartphone, a computer processing device, non- transitory computer readable medium, or alternately a computing device, microprocessor, or other computer type device independent of or incorporated with the present invention. An independent computing device can be networked together with the device either with wires or wirelessly. Indeed, any suitable method of analysis known to or conceivable by one of skill in the art could be used. It should also be noted that while specific equations are detailed herein, variations on these equations can also be derived, and this application includes any such equation known to or conceivable by one of skill in the art. [0032] A non-transitory computer readable medium is understood to mean any article of manufacture that can be read by a computer. Such non-transitory computer readable media includes, but is not limited to, magnetic media, such as a floppy disk, flexible disk, hard disk, reel-to-reel tape, cartridge tape, cassette tape or cards, optical media such as CD-ROM, writable compact disc, magneto-optical media in disc, tape or card form, and paper media, such as punched cards and paper tape. PATENT Page 11 P18228_02 [0033] Although the present invention has been described in connection with preferred embodiments thereof, it will be appreciated by those skilled in the art that additions, deletions, modifications, and substitutions not specifically described may be made without departing from the spirit and scope of the invention as defined in the appended claims.

Claims

PATENT Page 12 P18228_02 CLAIMS 1. A system comprising: an oximetry module wherein the oximetry module comprises a light source and wherein the oximetry module is configured to receive light from the light source after it is passed through a sample; and, a microprocessor configured to receive data from the oximetry module, wherein the microprocessor is configured to determine characteristics about the sample. 2. The system of claim 1 further comprising a sample well. 3. The system of claim 1 further comprising four oximetry modules. 4. The system of claim 2 further comprising four sample wells disposed in a sample tray. 5. The system of claim 1, wherein the system is configured for detecting a target analyte. 6. The system of claim 5 wherein the target analyte takes the form of a target biomolecule. 7. The system of claim 5, wherein the system is configured for detecting more than one target analyte simultaneously. 8. The system of claim 1, wherein the system is configured for detection of vascular endothelial growth factor (VEGF), angiopoietin-2 (Ang-2), monocyte chemoattractant protein-1 (MCP-1), and interleukin-6 (IL-6). 9. The system of claim 2 further comprising a number of oximetry modules corresponding to a number of sample wells. 10. The system of claim 1 wherein the microprocessor further comprises a display. 11. The system of claim 1 further comprising a power source. 12. The system of claim 11 wherein the power source comprises a battery. PATENT Page 13 P18228_02 13. The system of claim 1 wherein the microprocessor is configured to transmit data wirelessly. 14. A method for detecting characteristics of a sample comprising: applying light to the sample, wherein the light is a predetermined wavelength; detecting the light passed through the sample with an oximetry module; processing data from the oximetry module with a microprocessor to determine characteristics of the sample. 15. The method of claim 14 further comprising displaying data about the sample on a display of the microprocessor. 16. The method of claim 14 further comprising transmitting data from the microprocessor. 17. The method of claim 16 further comprising wirelessly transmitting the data. 18. The method of claim 14 further comprising detecting a presence of a target analyte. 19. The method of claim 14 further comprising detecting a concentration of a target analyte. 20. The method of claim 19 further comprising detecting the presence of a target biomolecule.