DESC:FIELD OF THE INVENTION
[0001] The present disclosure relates generally to the field of biochemistry. Specifically, the present disclosure relates to a kit, a device and an in-vitro method for screening a biomarker for assessing health risks.

BACKGROUND
[0002] Background description includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art.
[0003] Wellness is more than the absence of disease; it is a state of complete physical, mental and social well being. Life nowadays is generally much more stressful than in the past. Stress is the most common issue today, giving rise to lifestyle disorders and leading to compromise in wellness. Increased consumption of unhealthy fast food, travelling, financial pressure or emotional issues have further contributed towards this problem. All these factors may lead to disorders like chronic inflammation, tissue damage and low immunity.
[0004] Usually, underlying tissue inflammation and damage, which may be a result of untreated, undertreated infection/autoimmune/emotional stress/physical conditions along with long term exposure to irritants, such as industrial chemicals or polluted air, eventually start damaging healthy cells, tissues, and organs. This may lead to DNA damage, tissue death, and internal scarring. Symptoms present during this phase are usually subtler like mild fatigue, fever, mouth sores, rashes, abdominal pain, chest pain and are easy to overlook. This may negatively impact tissues and organs and ultimately lead to development of conditions like asthma, diabetes, neurological disorders or even cancer.
[0005] However, during this phase, body may show changes in levels of some signature biomolecules (biomarker), which may give some hint before the development of disease. A biomarker is a measurable indicator of the severity or presence of some disease state or some other physiological state in an organism. Biomarkers are molecules that indicate normal or abnormal processes taking place in our body and may act as molecular signatures for state of wellness. Various types of molecules, such as DNA, proteins or hormones, can serve as biomarkers, since they all indicate something about our health. Small changes in these biomarker levels may be indicative of big changes in health status. They reflect specific aspects of health and wellness based on diet, exercise and health status. Biomarkers can be: Predictive (early detection based on early stages of symptoms), Prognostic (disease management based on symptoms), or Diagnostic (presence or absence of a given disease).
[0006] Biomarkers are key factors that play crucial role in the development of diseases at cellular and molecular level. These signature molecules may help us identify the state of progression towards disease much before its actual manifestation. Timely detection and appropriate intervention may prevent or delay the disease and maintain the state of well-being.
[0007] There is therefore an unmet need in the art for identification and screening of such biomarkers that are indicative of the overall health and wellness status of a body. The present invention satisfies the existing as well as others needs.

OBJECTS
[0008] An object of the present disclosure is to provide a kit for screening a biomarker for assessing health risks.
[0009] Another object of the present disclosure is to provide a device for screening a biomarker for assessing health risks.
[0010] Another object of the present disclosure is to provide an in-vitro method for screening a biomarker for assessing health risks.
[0011] Another object of the present disclosure is to provide a kit, a device and a method for screening a biomarker that is rapid and accurate.
[0012] Another object of the present disclosure is to provide a kit, a device and a method for screening a biomarker that is sensitive.

SUMMARY
[0013] The present disclosure relates generally to the field of biochemistry. Specifically, the present disclosure relates to a kit, a device and an in-vitro method for screening a biomarker for assessing health risks.
[0014] An aspect of the present disclosure relates to a kit for screening a biomarker in a sample, said kit comprising: a support for placing the sample; a detecting agent; and a mean for multi-analyte profiling to detect and quantify the biomarker in the sample, wherein, the biomarker comprises a protein biomarker selected from the group comprising of ICAM-1, beta-NGF, GDNF, MPO, Park7/DJ-1, S100B, alpha-fetaprotein (AFP), FGF-23, Angiopoietin-like protein 3, adiponectin, beta-2-microglobulin, calbindin D, cystatin C, TIM-1/KIM-1, Dkk-1, osteopontin (OPN), osteoprotegerin (OPG), CD40, D-dimer, galectin-3, cardiac myoglobin, L-selectin, angiogenin, C-peptide, Leptin, insulin, glucagon, TNF-a, IFN-?, C-reactive protein (CRP), IL-8 and combinations thereof. In an embodiment, the support for holding the sample is selected from the group consisting of a micro-titer plate, a chip, a stick, a bead, a microbead, a glass and a membrane. In an embodiment, the detecting agent is selected from any or a combination of primary and secondary antibodies such as but not limited to Biotin-Antibody Cocktail, Streptavidin-PE and the like.
[0015] Another aspect of the present disclosure relates to a device for screening a biomarker in a sample, said device comprising: a support for placing the sample; and a mean for multi-analyte profiling to detect and quantify the biomarker in the sample, wherein, the biomarker comprises a protein biomarker selected from the group comprising of ICAM-1, beta-NGF, GDNF, MPO, Park7/DJ-1, S100B, alpha-fetaprotein (AFP), FGF-23, Angiopoietin-like protein 3, adiponectin, beta-2-microglobulin, calbindin D, cystatin C, TIM-1/KIM-1, Dkk-1, osteopontin (OPN), osteoprotegerin (OPG), CD40, D-dimer, galectin-3, cardiac myoglobin, L-selectin, angiogenin, C-peptide, Leptin, insulin, glucagon, TNF-a, IFN-?, C-reactive protein (CRP), IL-8 and combinations thereof. In an embodiment, the support for placing the sample is selected from the group consisting of a micro-titer plate, a chip, a stick, a bead, a microbead, a glass and a membrane.
[0016] Still another aspect of the present disclosure relates to an in vitro method of screening a biomarker in a sample, wherein the method comprises the steps of: placing the sample having biomarker on a support; adding a detecting agent into the sample having biomarker, wherein the detecting agent forms an immunological complex with the biomarker present in the sample; detecting and quantifying the biomarker by using a mean for multi-analyte profiling that identifies the immunological complexes to give a biomarker information set; and comparing the biomarker information set with a reference biomarker information set, wherein, the biomarker comprises a protein biomarker selected from the group comprising of ICAM-1, beta-NGF, GDNF, MPO, Park7/DJ-1, S100B, alpha-fetaprotein (AFP), FGF-23, Angiopoietin-like protein 3, adiponectin, beta-2-microglobulin, calbindin D, cystatin C, TIM-1/KIM-1, Dkk-1, osteopontin (OPN), osteoprotegerin (OPG), CD40, D-dimer, galectin-3, cardiac myoglobin, L-selectin, angiogenin, C-peptide, Leptin, insulin, glucagon, TNF-a, IFN-?, C-reactive protein (CRP), IL-8 and combinations thereof. In an embodiment, the support for placing the sample is selected from the group consisting of a micro-titer plate, a chip, a stick, a bead, a microbead, a glass and a membrane. In an embodiment, the comparison of the biomarker information set and the reference biomarker information set generate a differential numerical value assessing the health of a subject.
[0017] Other aspects, advantages, and salient features of the invention will become apparent to those skilled in the art from the following detailed description, which, taken in conjunction with the exemplary embodiments of the invention.

DETAILED DESCRIPTION
[0018] The following is a detailed description of embodiments of the disclosure. The embodiments are in such detail as to clearly communicate the disclosure. However, the amount of detail offered is not intended to limit the anticipated variations of embodiments; on the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the present disclosure as defined by the appended claims.
[0019] All publications herein are incorporated by reference to the same extent as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. Where a definition or use of a term in an incorporated reference is inconsistent or contrary to the definition of that term provided herein, the definition of that term provided herein applies and the definition of that term in the reference does not apply.
[0020] Reference throughout this specification to “one embodiment” or “an embodiment” means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment. Thus, the appearances of the phrases “in one embodiment” or “in an embodiment” in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.
[0021] In some embodiments, numbers have been used for quantifying weights, percentages, ratios, and so forth, to describe and claim certain embodiments of the invention and are to be understood as being modified in some instances by the term “about.” Accordingly, in some embodiments, the numerical parameters set forth in the written description and attached claims are approximations that can vary depending upon the desired properties sought to be obtained by a particular embodiment. In some embodiments, the numerical parameters should be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of some embodiments of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as practicable. The numerical values presented in some embodiments of the invention may contain certain errors necessarily resulting from the standard deviation found in their respective testing measurements.
[0022] Various terms as used herein are shown below. To the extent a term used in a claim is not defined below, it should be given the broadest definition persons in the pertinent art have given that term as reflected in printed publications and issued patents at the time of filing.
[0023] As used in the description herein and throughout the claims that follow, the meaning of “a,” “an,” and “the” includes plural reference unless the context clearly dictates otherwise. Also, as used in the description herein, the meaning of “in” includes “in” and “on” unless the context clearly dictates otherwise.
[0024] Unless the context requires otherwise, throughout the specification which follow, the word “comprise” and variations thereof, such as, “comprises” and “comprising” are to be construed in an open, inclusive sense that is as “including, but not limited to.”
[0025] The recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it were individually recited herein.
[0026] All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g. “such as”) provided with respect to certain embodiments herein is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention.
[0027] Groupings of alternative elements or embodiments of the invention disclosed herein are not to be construed as limitations. Each group member can be referred to and claimed individually or in any combination with other members of the group or other elements found herein. One or more members of a group can be included in, or deleted from, a group for reasons of convenience and/or patentability. When any such inclusion or deletion occurs, the specification is herein deemed to contain the group as modified.
[0028] The description that follows, and the embodiments described therein, is provided by way of illustration of an example, or examples, of particular embodiments of the principles and aspects of the present disclosure. These examples are provided for the purposes of explanation, and not of limitation, of those principles and of the disclosure.
[0029] It should also be appreciated that the present disclosure can be implemented in numerous ways, including as a system, a method or a device. In this specification, these implementations, or any other form that the invention may take, may be referred to as processes. In general, the order of the steps of the disclosed processes may be altered within the scope of the invention.
[0030] The headings and abstract of the invention provided herein are for convenience only and do not interpret the scope or meaning of the embodiments.
[0031] The following discussion provides many example embodiments of the inventive subject matter. Although each embodiment represents a single combination of inventive elements, the inventive subject matter is considered to include all possible combinations of the disclosed elements. Thus if one embodiment comprises elements A, B, and C, and a second embodiment comprises elements B and D, then the inventive subject matter is also considered to include other remaining combinations of A, B, C, or D, even if not explicitly disclosed.
[0032] The present disclosure relates generally to the field of biochemistry. Specifically, the present disclosure relates to a kit, a device and an in-vitro method for screening a biomarker for assessing health risks.
[0033] An aspect of the present disclosure relates to a kit for screening a biomarker in a sample, said kit comprising: a support for placing the sample; a detecting agent; and a mean for multi-analyte profiling to detect and quantify the biomarker in the sample, wherein, the biomarker comprises a protein biomarker selected from the group comprising of ICAM-1, beta-NGF, GDNF, MPO, Park7/DJ-1, S100B, alpha-fetaprotein (AFP), FGF-23, Angiopoietin-like protein 3, adiponectin, beta-2-microglobulin, calbindin D, cystatin C, TIM-1/KIM-1, Dkk-1, osteopontin (OPN), osteoprotegerin (OPG), CD40, D-dimer, galectin-3, cardiac myoglobin, L-selectin, angiogenin, C-peptide, Leptin, insulin, glucagon, TNF-a, IFN-?, C-reactive protein (CRP), IL-8 and combinations thereof.
[0034] In an embodiment, the support for holding the sample is selected from the group consisting of a micro-titer plate, a chip, a stick, a bead, a microbead, a glass and a membrane. Preferably, the micro-titer plate is a 96-well reaction plate. In an embodiment, the detecting agent is selected from any or a combination of primary and secondary antibodies such as but not limited to Biotin-Antibody Cocktail, Streptavidin-PE and the likes. However, a person skilled in the art would appreciate that any other detecting agent(s) or a combination of detecting agent(s) can be utilized to serve the intended purpose. In an embodiment, the mean for multi-analyte profiling comprise of a multiplex protein biomarker machine or a multiplex protein biomarker assays. More preferably, the means for multi-analyte profiling is a multiplex protein biomarker machine. In an embodiment, the immunological complexes may be detected by spectroscopic techniques including, but not limited to luminescence, fluorescence, absorbance, reflectance, electrochemical methods or combination thereof. In an embodiment, the said mean is capable of measuring multiple protein biomarkers sequentially or simultaneously. In an embodiment, the said mean is capable of measuring 1 to 50 biomarkers at a time. Most preferably, the levels of protein biomarkers are determined by Luminex Magpix™ technology that is capable of measuring up to 29 biomarkers. In an embodiment, the protein biomarkers may include the modified forms or isoforms of the protein biomarkers.
[0035] In an embodiment, the kit may further comprise an instruction sheet for suitable operation instructions.
[0036] Another aspect of the present disclosure relates to a device for screening a biomarker in a sample, said device comprising: a support for placing the sample; and a mean for multi-analyte profiling to detect and quantify the biomarker in the sample, wherein, the biomarker comprises a protein biomarker selected from the group comprising of ICAM-1, beta-NGF, GDNF, MPO, Park7/DJ-1, S100B, alpha-fetaprotein (AFP), FGF-23, Angiopoietin-like protein 3, adiponectin, beta-2-microglobulin, calbindin D, cystatin C, TIM-1/KIM-1, Dkk-1, osteopontin (OPN), osteoprotegerin (OPG), CD40, D-dimer, galectin-3, cardiac myoglobin, L-selectin, angiogenin, C-peptide, Leptin, insulin, glucagon, TNF-a, IFN-?, C-reactive protein (CRP), IL-8 and combinations thereof. In an embodiment, the support for placing the sample is selected from the group consisting of a micro-titer plate, a chip, a stick, a bead, a microbead, a glass and a membrane. Preferably, the micro-titer plate is a 96-well reaction plate.
[0037] Still another aspect of the present disclosure relates to an in vitro method of screening a biomarker in a sample, wherein the method comprises the steps of: placing the sample having biomarker on a support; adding a detecting agent into the sample having biomarker, wherein the detecting agent forms an immunological complex with the biomarker present in the sample; detecting and quantifying the biomarker by using a mean for multi-analyte profiling that identifies the immunological complexes to give a biomarker information set; and comparing the biomarker information set with a reference biomarker information set, wherein, the biomarker comprises a protein biomarker selected from the group comprising of ICAM-1, beta-NGF, GDNF, MPO, Park7/DJ-1, S100B, alpha-fetaprotein (AFP), FGF-23, Angiopoietin-like protein 3, adiponectin, beta-2-microglobulin, calbindin D, cystatin C, TIM-1/KIM-1, Dkk-1, osteopontin (OPN), osteoprotegerin (OPG), CD40, D-dimer, galectin-3, cardiac myoglobin, L-selectin, angiogenin, C-peptide, Leptin, insulin, glucagon, TNF-a, IFN-?, C-reactive protein (CRP), IL-8 and combinations thereof.
[0038] In an embodiment, the biomarker comprises protein biomarkers selected from the group comprising of ICAM-1, beta-NGF, GDNF, MPO, Park7, S100B, or combinations thereof. In AN embodiment, a differential value of the biomarker information set and the reference biomarker information set maybe indicative of a risk to brain. Risk to brain health may be caused due to depression, anxiety, stress, mood disorders and the likes.
[0039] In an embodiment, the biomarker comprises protein biomarkers selected from the group comprising of ICAM-1, MPO, Leptin, insulin, C-peptide, FGF-23, Angiopoietin-like protein 3, adiponectin, C-reactive protein, galectin-3, or combinations thereof. In an embodiment, a differential value of the biomarker information set and the reference biomarker information set maybe indicative of a risk to metabolic health. Risk to metabolic health may be caused due to diabetes, hypertension, obesity, hypercholesterolemia, insulin resistance, kidney disorders, atherosclerosis, fatty liver disease and the likes.
[0040] In an embodiment, the biomarker comprises protein biomarkers selected from the group comprising of CD-40, D-dimer, cardiac myoglobin, L-selectin, Angiopoietin-like protein 3, angiogenin, galectin-3, or combinations thereof. In an embodiment, a differential value of the biomarker information set and the reference biomarker information set maybe indicative of a risk to cardiovascular health. Risk to cardiovascular health may be caused due to hypertension, thrombosis, cardiac fibrosis, myocardial infarction, atherosclerosis and the likes.
[0041] In an embodiment, the biomarker comprises protein biomarkers selected from the group comprising of TNF-a, IFN-?, C-reactive protein (CRP), IL-8, or combinations thereof. In an embodiment, a differential value of the biomarker information set and the reference biomarker information set maybe indicative of an inflammation. Inflammation may be a possible indication of infections, allergic diseases, chronic stress, inflammatory bowel disease, arthritis, diabetes, sepsis, atherosclerosis and the likes.
[0042] In an embodiment, the biomarker panel comprises protein biomarkers selected from the group comprising of TNF-a, IFN-?, C-reactive protein (CRP), IL-8, or combinations thereof. In an embodiment, a differential value of the biomarker information set and the reference biomarker information set maybe indicative of an inflammation.
[0043] In an embodiment, the support for placing the sample is selected from the group consisting of a micro-titer plate, a chip, a stick, a bead, a microbead, a glass and a membrane. Preferably, the micro-titer plate is a 96-well reaction plate. In an embodiment, the comparison of the biomarker information set and the reference biomarker information set generate a differential numerical value assessing the health of a subject. The panel information set comprises the values of all the protein biomarkers in the sample. The biomarker information set may optionally be plotted into biomarker level graphs for ease of assessment.
[0044] In an embodiment, the reference biomarker information set may comprise of threshold values for each individual biomarker. The skilled artisan should understand that numerous methods may be used to select the threshold values for a particular biomarker or a plurality of biomarkers. In an embodiment, the comparison of the biomarker information set and the reference biomarker information set reveals the up-regulation or down-regulation of the particular protein biomarker. Data generated thereafter can be interpreted to assess the health risks to a subject.
[0045] In an embodiment, the method may be run in iterations to cover the entire range of protein biomarkers. In an embodiment, a study of the values of the protein biomarkers in a sample of the subject may be used to indicate a health risk.
[0046] In an embodiment, an appropriate amount of the sample may be obtained from cells or tissues of the subject and it may be any body fluid, including but not limited to blood, plasma, urine, saliva, cerebral fluid, or serum. In a preferable embodiment, the sample is blood or serum.
[0047] In an embodiment, the appropriate amount of the sample may be about 2 ml to up to 100 ml. The concentration of the protein biomarker in the sample that is detectable may be as low as the picograms/milliliter range.
[0048] In an embodiment, the present disclosure is capable of providing a rapid and reliant analysis of the sample. In an embodiment, the present disclosure reduces the costs of healthcare.

ADVANTAGES
[0049] The present disclosure provides a kit for screening a biomarker for assessing health risks.
[0050] The present disclosure provides a device for screening a biomarker for assessing health risks.
[0051] The present disclosure provides an in-vitro method for screening a biomarker for assessing health risks.
[0052] The present disclosure provides a kit, a device and a method for screening a biomarker that is rapid and accurate.
[0053] The present disclosure provides a kit, a device and a method for screening a biomarker that is sensitive.

WE CLAIMS:

1. A kit for screening a biomarker in a sample, said kit comprising:
a support for placing the sample;
a detecting agent; and
a mean for multi-analyte profiling to detect and quantify the biomarker in the sample,
wherein, the biomarker comprises a protein biomarker selected from the group comprising of ICAM-1, beta-NGF, GDNF, MPO, Park7/DJ-1, S100B, alpha-fetaprotein (AFP), FGF-23, Angiopoietin-like protein 3, adiponectin, beta-2-microglobulin, calbindin D, cystatin C, TIM-1/KIM-1, Dkk-1, osteopontin (OPN), osteoprotegerin (OPG), CD40, D-dimer, galectin-3, cardiac myoglobin, L-selectin, angiogenin, C-peptide, Leptin, insulin, glucagon, TNF-a, IFN-?, C-reactive protein (CRP), IL-8 and combinations thereof.

2. The kit as claimed in claim 1, wherein the support for holding the sample is selected from the group consisting of a micro-titer plate, a chip, a stick, a bead, a microbead, a glass and a membrane.

3. A device for screening a biomarker in a sample, said device comprising:
a support for placing the sample; and
a mean for multi-analyte profiling to detect and quantify the biomarker in the sample,
wherein, the biomarker comprises a protein biomarker selected from the group comprising of ICAM-1, beta-NGF, GDNF, MPO, Park7/DJ-1, S100B, alpha-fetaprotein (AFP), FGF-23, Angiopoietin-like protein 3, adiponectin, beta-2-microglobulin, calbindin D, cystatin C, TIM-1/KIM-1, Dkk-1, osteopontin (OPN), osteoprotegerin (OPG), CD40, D-dimer, galectin-3, cardiac myoglobin, L-selectin, angiogenin, C-peptide, Leptin, insulin, glucagon, TNF-a, IFN-?, C-reactive protein (CRP), IL-8 and combinations thereof.

4. The device as claimed in claim 3, wherein the support for placing the sample is selected from the group consisting of a micro-titer plate, a chip, a stick, a bead, a microbead, a glass and a membrane.

5. An in vitro method of screening a biomarker in a sample, wherein the method comprises the steps of:
(a) placing the sample having biomarker on a support;
(b) adding a detecting agent into the sample having biomarker, wherein the detecting agent forms an immunological complex with the biomarker present in the sample;
(c) detecting and quantifying the biomarker by using a mean for multi-analyte profiling that identifies the immunological complexes to give a biomarker information set; and
(c) comparing the biomarker information set with a reference biomarker information set,
wherein, the biomarker comprises a protein biomarker selected from the group comprising of ICAM-1, beta-NGF, GDNF, MPO, Park7/DJ-1, S100B, alpha-fetaprotein (AFP), FGF-23, Angiopoietin-like protein 3, adiponectin, beta-2-microglobulin, calbindin D, cystatin C, TIM-1/KIM-1, Dkk-1, osteopontin (OPN), osteoprotegerin (OPG), CD40, D-dimer, galectin-3, cardiac myoglobin, L-selectin, angiogenin, C-peptide, Leptin, insulin, glucagon, TNF-a, IFN-?, C-reactive protein (CRP), IL-8 and combinations thereof.

6. The method as claimed in claim 5, wherein the support for placing the sample is selected from the group consisting of a micro-titer plate, a chip, a stick, a bead, a microbead, a glass and a membrane.
7. The method as claimed in claim 5, wherein the comparison of the biomarker information set and the reference biomarker information set generate a differential numerical value assessing the health of a subject.