Improved methods for determining cardiac myosin binding protein c in biological samples

Abstract

Provided herein are improved assays, methods, and systems for determining the amount, concentration, or level of cardiac myosin binding protein C ("cMyC") in a biological sample obtained from a subject. The improved assays, methods, and systems can generate a result (e.g., the amount or level of cMyC) in less than about 1 hour and are highly specific and sensitive.

Description

[0001] Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 IMPROVED METHODS FOR DETERMINGCARDIAC MYOSIN BINDING PROTEIN C IN BIOLOGICAL SAMPLES Related Application Information This application claims the benefit of U.S. Provisional Application No.63 / 641,083, filed May 1, 2024, the content of which is herein incorporated by reference in its entirety. Sequence Listing Statement The contents of the electronic sequence listing titled ABBTL_43031_601_SequenceListing.xml (Size: 5,484bytes; and Date of Creation: April 28, 2025) is herein incorporated by reference in its entirety. Technical Field The present disclosure relates to improved assays, methods, and systems for determining the amount, concentration,or level of cardiac myosin binding protein C (“cMyC”) in a biological sample obtained from a subject. In some embodiments, the subject has or is suspected of having a cardiac injury. Additionaly, the improved assays, methods, and systems of the present disclosure can generatea result (e.g., the amount or level of cMyC) in less than one hourfrom the start of the assay or method and are highly specific and sensitive. Background Chest pain isthe predominant symptom of acute myocardial infarction (AMI), commonly known as a heart atack. When a patient presents with suspected AMI, triage heavily relies on a range of measures, including conducting electrocardiography(EKG) andassessing cardiac Troponin (cTn) levels. EKGsare diagnostic teststhatrecord the heart's electrical activity typicaly obtained by placing electrodes on the skin. However, EKG may not always detect AMI, particularly in cases of atypical presentation or when heart damage is insuficient to produce characteristic electrical changes. Additionaly, technical issues in recording or interpreting EKG and concurent medical conditions can impact their accuracy in diagnosing AMI. Guidelines indicate that healthcare providers should perform anassessment of cTn levels to evaluate the risk of AMIin al suspected cases(Gulati et al., Cardiovasc Comput Tomogr., Jan-Feb;16(1):54-122 (2022).Elevated cTn levels indicate myocardial damage that may be associated with conditions Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 like AMI. However, one significant drawback is that cTn is released into the bloodstream relatively gradualy and clears slowly. As a result, patients may needmultiple blood tests over time to monitor changes in cTn levels accurately. Moreover, some individuals might consistently display elevated cTn levels due to factors such as chronic heart disease or other underlying conditions, complicating the interpretation of results and posing a chalenge in maintaining an optimal balance between sensitivity and specificity in AMI diagnosis. Delays in the development of more accurate and rapiddiagnostic tools for MInot only results in financial burdenand medical strife for the patient but, also has significant connotations for the patient’s overalpsychological, and social wel-being as wel as creates a burden and liability on the healthcare systems. For example, from a medical perspective, delays in diagnosis can exacerbate the severity of the heart atack, leading to increased morbidity and mortality rates. Additionaly, the uncertainty surounding the diagnosis may contribute to heightened anxiety and stress levels among patients, negatively impacting their overal wel-being and quality of life. Furthermore, prolonged diagnostic processes can strain healthcare resources, leading to increased healthcare costs and potential ineficiencies in the delivery of care. On a broader societal level, the repercussions of delayed AMI diagnosis may extend to reduced workforce productivity, increased absenteeism, and societal burdens associated with long-term disability and rehabilitation. Therefore, swift, and accurate diagnosis of AMI is crucial not only for mitigating the immediate health risks but also for aleviating the broader economic and societal burdens imposed by this condition. Given these findings,the development of new biomarkers is imperative to address the chalenges inherent in curent diagnostic methods. To surpass high sensitivity cTn immunoassays as the gold standard, future biomarkers must not only demonstrate heightened sensitivity and selectivity in assessing the risk of AMI or other cardiac-related conditions but also exhibit superior characteristics compared to cTn immunoassays. This entails maintaining equivalent cardiac selectivity while showcasing a more rapid rise folowing acute myocardial injury, thereby enhancing sensitivity. Moreover, these biomarkers should idealy present with lower 'background' concentrations in individuals predisposed to vascular risk factors or underlying chronic heart disease, thereby refining specificity in diagnosis. The ideal biomarker for early AMI detection would rise rapidly and abundantly at the earliest stages of myocardial necrosis,while embodying the cardiac-restricted expression halmark of cTns, thus ensuring precise and timely diagnosis. Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 Consequently, there is an urgent need for the development of highly sensitive and specific immunoassay tests capable ofmore rapidly andaccurately diagnosing acute myocardial infarction (AMI) and other chronic heart conditions, which would significantly improve patient outcomes and healthcare delivery. SUMMARY OF THE INVENTION Provided herein is a method comprising: a) receiving a level of cardiac myosin binding protein C (“cMyC”) from an assay performed on at least one biological sample obtained from a subject; and b) determining whether the level of cMyC obtained in step a) is higher or lower than a reference level, wherein: i. if the level of cMyC determined in step b) is: (A) higher than the reference level, further determining that the subject is sufering from a cardiac injury; or (B) lower than the reference level, further determining that the subject is not sufering from acardiac injury; and i. performing the assay comprises: A)contacting the biological sample either simultaneously or sequentialy, in any order, with: at least one first specific binding partner atached to a solid support, wherein the at least one first specific binding partner comprises monoclonal antibody 3H8 which specificaly binds to at least one epitope on cMyC, and at least one second specific binding partner comprising a detectable label, wherein the at least one second specific binding partner comprises monoclonal antibody 1A4 which specificaly binds to at least one epitope on cMyC which is a diferent epitope than the epitope bound by monoclonal antibody 3H8, to form a first mixture, B) incubating the first mixture for about 12 to about 30 minutes to form one or more first complexes comprising the first specific binding partner-cMyC-second specific binding partner; C) isolating the first specific binding partner-cMyC-second specific binding partner complexes from the first mixture; Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 D) washing the first specific binding partner-cMyC-second specific binding partner complex of step C) with a solution comprising water or at least one detergent to form a second mixture comprising the first specific binding partner-cMyC-second specific binding partner; E) incubating the second mixture comprising the first specific binding partner-cMyC- second specific binding partner complex for about 1 to about 5 minutes; F) isolating the first specific binding partner-cMyC-second specific binding partner complexes from the second mixture; G) washing the isolated first specific binding partner-cMyC-second specific binding partner complex of step F) with a solution comprising water or at least one detergent; and H) assessing a signal from the one or more first complexes, wherein the amount of detectable signal from the detectable label indicates the presence or amount of cMyC in the sample. In some embodiments, the biological sample is a whole blood, serum, or plasma sample. In some embodiments, the biological sample is a whole blood sample. In some embodiments, the biological sample is a serum sample. In some embodiments, the biological sample is a plasma sample. In some embodiments, the solid support is a microparticle. In some embodiments, the cardiac injury myocardial infarction or reinfarction. In some embodiments, the antibody 3H8 is produced by hybridoma cel lineDSM ACC3223. In some embodiments, the antibody 1A4 is produced by hybridoma cel line DSM ACC3224. In some embodiments, the method comprises determining that the subject is sufering from a cardiac injury. In some embodiments, the cardiac injury is myocardial infarction or reinfarction. In some embodiments, the myocardial infarction is acute myocardial infarction. In some embodiments, the method further comprises treating the patient for the cardiac injury, wherein the treatment comprises: (a) administering to the subject one or more pharmaceutical agents, oxygen, or a combination thereof; (b) performing one or more procedures Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 on the subject, wherein said procedures are angioplasty, inserting a stent, atherectomy, bypass surgery, or combinations thereof; or (c) any combination of (a) and (b). In some embodiments, the method further comprises monitoring a subject sufering a cardiac injury. In some further embodiments, the assay is an immunoassay or a clinical chemistry assay. In some further embodiments, the assay is performed using single molecule detection, lateral flow, or a point-of care method. In yet further embodiments, the first mixture is incubated for about 18 minutes. In yet further embodiments, the second mixture is incubated for about 4 minutes. In yet further embodiments, the method is quantitative. In stil further embodiments, the method is qualitative. BRIEF DESCRIPTION OF THE DRAWINGS Having thus described the presently disclosed subject mater in general terms, reference wil now be made to the accompanying Figures, which are not necessarily drawn to scale, and wherein: FIG.1: A schematic of themethods described in Example 1. FIG.2A-B: (A) A schematic representation of cMyC (yelow; Cardiac Specific motifs) and (B)shows sequences of SEQ ID NOS.: 1 and 2 which include epitopes 3H8 and 1A4 (in bold). FIG.3: Demonstrates cMyC concentration (ng / L) versus percent (%) CV as based on Example 1. FIG.4:Shows expected versus observed cMyC concentration (ng / L) based on Example 1. FIG.5: Shows the detection of cMyCin samples obtained from a normal, healthy population as described in Example 1. FIG.6: Shows the detection of cMyC in samples obtained from subjects having elevated cTnI levels as described in Example 1. Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 DETAILED DESCRIPTION Provided herein are improved assays, methods, and systemsfor determining the amount, concentration, or levelof cardiac myosin binding protein C (“cMyC”)in at least one biological sample obtained from a subject. In some embodiments, the assays, methods, and systems described herein can be used to determine whether a subject is suffering from or has suffered at least onecardiac injury. In some embodiments, the assays, methods, and systemsof the present disclosure employ a unique combination ofantibodiesas capture and detection antibodies. Specificaly, the assays, methods, and systems of the present disclosureutilize monoclonal antibody 3H8 as a capture antibody (e.g.,first specific binding partner)and monoclonal antibody 1A4, labeled with at least one detectable label, as a detection antibody(e.g., a second specific binding partner). In addition to the unique combination of capture and detection antibodies, the assays, methods, and systems also employ certainincubation and wash steps. As a result, the assays,methods, and systemsof the present disclosure exhibit greater analytical sensitivity, namely,about a 3-fold lower limit of detection and about a 2-fold lower limit of quantification,when compared tothe prior art methods, specificaly, the cMyC assay for use on theErenna platform (Singulex). The cMyC assay for use on the Erenna platform utilizes monoclonal antibody 1A4as the capture antibody and antibody 3H8 labeled with a detectable label, as the detection antibody.Additionaly, as discussed in Marjot, J., “The development and application of a high-sensitivity immunoassay for cardiac myosin-binding protein C,” Trans. Res., 170:17-25.eg(April 2016), the cMyC assay optimized for use on the Erenna platform takes generates a result more than 3 hours after the start of the assay. Specificaly,in the assay optimized for use on the Erenna platform, microparticles coated with monoclonal antibody 1A4are incubated with a sample for 2 hours. After a2-hour incubation, monoclonal antibody 3H8, labeled with a detectable label, is added to the reaction mixture and incubated for 1 hour and then washed the result determined using single molecule counting. In addition to the improved analytical sensitivity, the assays and methods of the present disclosure alsogenerate a result (e.g., the amount, level or concentration of cMyC) inlessone hour, and in some instances, lessthan about 30 minutes from the start of the assayor method. In contrast, the cMyC assay for use on theErenna platform generates a result in about 3hours Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 from the start of the assay(Marjot, J., “The development and application of a high-sensitivity immunoassay for cardiac myosin-binding protein C,” Trans. Res., 170:17-25.eg (April 2016)). DEFINITIONS The terms "comprise(s)," "include(s)," "having," "has," "can," "contain(s)," and variants thereof, as used herein, are intended to be open-ended transitional phrases, terms, or words that do not preclude the possibility of additional acts or structures. The singular forms "a," "and," and "the" include plural references unless the context clearly dictates otherwise. The present disclosure also contemplates other embodiments "comprising," "consisting of,"and "consisting essentialy of," the embodiments or elements presented herein, whether explicitly set forth or not. For the recitation of numeric ranges herein, each intervening number there between with the same degree of precision is explicitly contemplated. For example, for the range of 69, the numbers 7 and 8 are contemplated in addition to 6 and 9, and for the range 6.0-7.0, thenumber 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are explicitly contemplated. Unless otherwise defined herein, scientific, and technical terms used in connection with the present disclosure shal have the meanings that are commonly understood by those of ordinary skil in the art. The meaning and scope of the terms should be clear; in the event, however of any latent ambiguity, definitions provided herein take precedent over anydictionary or extrinsic definition. Further, unless otherwise required by context, singular terms shal include pluralities and plural terms shal include the singular. “3H8” or “monoclonal antibody 3H8” as used interchangeably herein, refers to the monoclonal antibody described in U.S. Patent No.9,975,961, the contents of which are herein incorporated by reference. Monoclonal antibody 3H8 specificaly binds to an epitope having the amino acid sequence of: APDDPIGLFVM (SEQ ID NO:1). In some embodiments, monoclonal antibody 3H8 can be produced by hybridoma cel line deposited with the DSMZ and assigned Accession Number DSM ACC3223. In other embodiments, monoclonal antibody 3H8 can be produced by recombinant means using recombinant DNA technology (e.g., the antibody is generated in vitrousing synthetic genes introduced into mammalian cel lines rather than through hybridoma cel culture or animal immunization), which is wel known in the art.In stil other embodiments, chimeric, CDR-grafted, humanized,fuly human, andafinity matured versions of 3H8 can be produced and used in the methods described herein using routine techniques known in the art. Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 “Antibody” and “antibodies” as used herein refers to monoclonal antibodies, multispecific antibodies, human antibodies, humanized antibodies (fuly or partialy humanized), animal antibodies such as, but not limited to, a bird (for example, a duck or a goose), a shark, a whale, and a mammal, including a non-primate (for example, a cow, a pig, a camel, a lama, a horse, a goat, a rabbit, a sheep, a hamster, a guinea pig, a cat, a dog, a rat, a mouse, etc.) or a non-human primate (for example, a monkey, a chimpanzee, etc.), recombinant antibodies, chimeric antibodies, single-chain Fvs (“scFv”), single chain antibodies, single domain antibodies, Fab fragments, F(ab') fragments, F(ab')2fragments, disulfide-linked Fvs (“sdFv”), and anti-idiotypic (“anti-Id”) antibodies, dual-domain antibodies, dual variable domain (DVD) or triple variable domain (TVD) antibodies (dual-variable domain immunoglobulins and methods for making them are described in Wu, C., et al., Nature Biotechnology, 25(11):1290-1297 (2007) and PCT International Application WO 2001 / 058956, the contents of each of which are herein incorporated by reference), and functionaly active epitope-binding fragments of any of the above. Antibodies include immunoglobulin molecules and immunologicaly active fragments of immunoglobulin molecules, namely, molecules that contain an analyte-binding site. Immunoglobulin molecules can be of any type (for example, IgG, IgE, IgM, IgD, IgA, and IgY), class (for example, IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass. For simplicity sake, an antibody against an analyte is frequently refered to herein as being either an “anti-analyte antibody” or merely an “analyte antibody” (e.g.,an anti-cMyCantibody or a cMyCantibody). An antibody as used herein also refers tonon-Ig derived alternatives (so-caled antibody ‘mimetics’) such as, e.g., aptamers, DARPins, Afimers, Avimers, Knotins, Monobodies, and Afinity Clamps. As used herein, the term “aptamer” refers to a nucleic acid that has a specific binding afinity for a target analyte or molecule. It is recognized that affinity interactions are a mater of degree; however, in this context, the “specific binding afinity” of anaptamerfor its analyte or target means that theaptamerbinds to its analyte or target generaly with a much higher degree of affinity than it binds to other components in a sample. “Antibody fragment” as used herein refers to a portion of an intact antibody comprising the antigen-binding site or variable region. The portion does not include the constant heavy chain domains (i.e.,CH2, CH3, or CH4, depending on the antibody isotype) of the Fc region of the intact antibody. Examples of antibody fragments include, but are not limited to, Fab fragments, Fab' fragments, Fab'-SH fragments, F(ab')2fragments, Fd fragments, Fv Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 fragments, diabodies, single-chain Fv (scFv) molecules, single-chain polypeptides containing only one light chain variable domain, single-chain polypeptides containing the three CDRs of the light-chain variable domain, single-chain polypeptides containing only one heavy chain variable region, and single-chain polypeptides containing the three CDRs of the heavy chain variable region. “Aptamer” as used hereinrefers to a nucleic acid that has a specific binding afinity for a target analyte or molecule. It is recognized that afinity interactions are a mater of degree; however, in this context, the “specific binding afinity” of an^aptamer^for its analyte or target means that the^aptamer^binds to its analyte or target generaly with a much higher degree of affinity than it binds to other components in a biological sample. An “aptamer” is a set of copies of one type or species of nucleic acid molecule that comprises a particular nucleotide sequence. An^aptamer^can include any suitable number of nucleotides, including any number of chemicaly modified nucleotides. “Aptamers” refers to more than one such set of molecules. Diferent^aptamers^can have either the sameor different numbers of nucleotides.^Aptamers^can be DNA or RNA or chemicaly modified nucleic acids and can be single-stranded, double-stranded, or contain double-stranded regions, and can include higher ordered structures. An^aptamer^can also be a photoaptamer, where a photoreactive or chemicaly reactive functional group is included in the^aptamer^to alow it to be covalently linked to its coresponding analyte or target. In some aspects, an^aptamer^may include a detectable label. “Bead” and “particle” are used herein interchangeably and refer to a substantialy spherical solid support. One example of a bead or particle is a microparticle. Microparticles that can be used herein can be any type known in the art. For example, the bead or particle can be a magnetic bead or magnetic particle. Magnetic beads / particles may be feromagnetic, ferimagnetic, paramagnetic, superparamagnetic or ferrofluidic. Exemplary feromagnetic materials include Fe, Co, Ni, Gd, Dy, CrO2, MnAs, MnBi, EuO, and NiO / Fe. Examples of ferimagnetic materials include NiFe2O4, CoFe2O4, Fe3O4(or FeO.Fe2O3).Beads can have a solid core portion that is magnetic and is surounded by one or more non-magnetic layers. Alternately, the magnetic portion can be a layer around a non-magnetic core. The microparticles can be of any size that would work in the methods described herein, e.g., from about 0.75 to about 5 nm, or from about 1 to about 5 nm, or from about 1 to about 3 nm. In other embodiments, the bead or Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 particle can be a non-magnetic bead or particle, such as for example, non-magnetic polystyrene particles. “Binding protein” is used herein to refer to a monomeric or multimeric protein that binds to and forms a complex with a binding partner, such as, for example, a polypeptide, an antigen, a chemical compound or other molecule, or a substrate of any kind. A binding protein specificaly binds a binding partner. Binding proteins include antibodies, as wel as antigen- binding fragments thereof and other various forms and derivatives thereof as are known in the art and described herein below, and other moleculescomprising one or more antigen-binding domains that bind to an antigen molecule or a particular site (epitope) on the antigen molecule. Accordingly, a binding protein includes, but is not limited to, an antibody a tetrameric immunoglobulin, an IgG molecule, an IgG1 molecule, a monoclonal antibody, a chimeric antibody, a CDR-grafted antibody, a humanized antibody, an afinity matured antibody, and fragments of any such antibodies that retain the ability to bind to an antigen. “Bispecific antibody” is used herein to refer to a ful-length antibody that is generated by quadroma technology (see Milstein et al.,Nature, 305(5934): 537-540 (1983), by chemical conjugation of two diferent monoclonal antibodies (see, Staerz et al.,Nature,314(6012): 628- 631 (1985), or by knob-into-hole or similar approaches, which introduce mutations in the Fc region (see Holiger et al.,Proc. Natl. Acad. Sci. USA, 90(14): 6444-6448 (1993), resulting in multiple diferent immunoglobulin species ofwhich only one is the functional bispecific antibody. A bispecific antibody binds one antigen (or epitope) on one of its two binding arms (one pair of HC / LC), and binds a diferent antigen (or epitope) on its second arm (a different pair of HC / LC). By this definition, a bispecific antibody has two distinct antigen-binding arms (in both specificity and CDR sequences), and is monovalent for each antigen to which it binds to. As used herein, the term "cardiac myosin-binding protein C” or “cMyC" refers to a cardiac biomarker known as cardiac myosin-binding protein C. It is involved in the regulation of cardiac muscle function and is associated with various cardiac-related events and conditions, such as cardiac injury.Exemplary adverse cardiac-related events and diseases in which cMyC may play a role include acute myocardial infarction, congestive heart failure, cardiomyopathies, arhythmias, and other conditions afecting the heart's structure and function. For example, elevated levels of cMyC in the bloodstream have been observed in patients folowing an acute myocardial infarction, indicating cardiac muscle damage. Additionaly, cMyC levels may be Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 monitored in individuals with congestive heart failure or adverse cardiac related conditions to assess disease severity and response to treatment. “Cardiac injury” or “myocardial injury”, as used interchangeably herein,refers to decreased blood flow through one or more coronary arteriesin a subject causing a reduction in the amount of oxygen being received by the muscle in the heart thereby resulting in damage to one or more myocardial cels. In some embodiments, cardiac or myocardial injury can be diagnosed or determined based on cardiac troponin (cTn) concentrationor levelsin a subject. According to the Fourth Universal Definition of Myocardial Infarction (2018)(Thygesen K., “Ten Commandments’ for the Fourth Universal Definition of Myocardial Infarction 2018”Eur Heart J., 40(3):226 (2019), a cardiac or myocardial injury is present when a subject exhibitsan elevation of cardiac troponin values with at least one value above the 99th percentile upper reference limit (URL) (often refered to as a “critical value”). Cardiac or myocardial injury can be considered acute when there is a rise and / or fal of troponin valuesin a subject, with at least one above the 99thpercentile.When chronic, the troponin levels in a subject remain stable at a constantly elevated level. In some embodiments, cardiac or myocardial injury may be due to ischemic or nonischemic causes.In some embodiments, cardiac or myocardial injury is due to one or more myocardial infarctions or reinfarctions. In some embodiments, the cardiac or myocardial injury is due to one or more myocardial infarctions. In another embodiment, the cardiac or myocardial injury is due to one or more reinfarctions. As used herein, the term “cardiac Troponin” refers to any of the subunits of the troponin complex: troponin I (TnI), troponin T (TnT) and troponin C (TnC). The troponin complex is located on the thin filament of the muscle contractile apparatus and plays an integral role in contraction of skeletal and cardiac muscle. The individual subunits of the complex have diferent functions: TnC binds to calcium ions to confer conformational change on TnI, which binds to actin in thin myofilaments to stabilize the actin-tropomyosin complex and TnTbinds to tropomyosin thereby forming a troponin-tropomyosin complex. Cardiac forms of TnIand TnTwere originaly used as markers for cardiac cel death, but both may be used to diagnose acute myocardial infarction, unstable angina, post-surgery myocardium trauma and some other related diseases with cardiac muscle injury (e.g., drug and toxin induced cardiomyocyte toxicity).In some embodiments, the cardiac troponin is cardiac troponinT (cTnT). In some embodiments, the cardiac troponin is cardiac troponinI (cTnI). Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 As used herein, the term "cardiovascular disease" and “adverse cardiac related events” refers to diseases, disorders and conditions of the heart and circulatory system. Exemplary cardiovascular diseases, including cholesterol-or lipid-related disorders, include, but are not limited to, acute coronary syndrome, angina, arteriosclerosis, atherosclerosis, carotid atherosclerosis, cerebrovascular disease, cerebral infarction, congestive heart failure, congenital heart disease, coronary heart disease, coronary artery disease, coronary plaque stabilization, dyslipidemias, dyslipoproteinemias, endothelium dysfunctions, familial hypercholesterolemia, familial combined hyperlipidemia, hypoalphalipoproteinemia, hypertriglyceridemia, hyperbetalipoproteinemia, hypercholesterolemia, hypertension, hyperlipidemia, intermitent claudication, ischemia, ischemia reperfusion injury, ischemic heart diseases, cardiac ischemia, metabolic syndrome, multi-infarct dementia, myocardial infarction, obesity, peripheral vascular disease, reperfusion injury, restenosis, renal artery atherosclerosis, rheumatic heart disease, stroke, thrombotic disorder, transitory ischemic atacks, and lipoprotein abnormalities associated with Alzheimer's disease, obesity, diabetes melitus, syndrome X, impotence, multiple sclerosis, Parkinson's disease, and inflammatory diseases. “CDR” is used herein to refer to the “complementarity determining region” within about an antibody variable sequence. There are three CDRs in each of the variable regions of the heavy chain and the light chain. Proceeding from the N-terminus of a heavy or light chain, these regions are denoted "CDR1", "CDR2", and "CDR3", for each of the variable regions. The term "CDR set" as used herein refers to a group of three CDRs that occur in a single variable region that binds the antigen. An antigen-binding site, therefore, may include six CDRs, comprising the CDR set from each of a heavy and a light chain variable region. A polypeptide comprising a single CDR, (e.g.,a CDR1, CDR2, or CDR3) may be refered to as a “molecular recognition unit.” Crystalographic analyses of antigen-antibody complexes have demonstrated that the amino acid residues of CDRs form extensive contact with bound antigen, wherein the most extensive antigen contact is with the heavy chain CDR3. Thus, the molecular recognition units may beprimarily responsible for the specificity of an antigen-binding site. In general, the CDR residues are directly and most substantialy involved in influencing antigen binding. The exact boundaries of these CDRs have been defined diferently according to diferent systems. The system described by Kabat (Kabat et al.,Sequences of Proteins of Immunological Interest(National Institutes of Health, Bethesda, Md. (1987) and (1991)) not Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 only provides an unambiguous residue numbering system applicable to any variable region of an antibody, but also provides precise residue boundaries defining the three CDRs. These CDRs may be referred to as "Kabat CDRs". Chothia and coworkers (Chothia and Lesk, J. Mol. Biol., 196: 901-917 (1987); and Chothia et al., Nature,342: 877-883 (1989) found that certain sub- portions within Kabat CDRs adopt nearly identical peptide backbone conformations, despite having great diversity at the level of amino acid sequence. These sub-portions were designated as "L1", "L2", and "L3", or "H1", "H2", and "H3", where the "L" and the "H" designate the light chain and the heavy chain regions, respectively. These regions may be refered to as "Chothia CDRs", which have boundaries that overlap with Kabat CDRs. Other boundaries defining CDRs overlapping with the Kabat CDRs have been described by Padlan, FASEB J., 9: 133-139 (1995), and MacCalum, J. Mol. Biol., 262(5): 732-745 (1996). Stil other CDR boundary definitions may not strictly folow one of the herein systems, but wil nonetheless overlap with the Kabat CDRs, although they may be shortened or lengthened in light of prediction or experimental findings that particular residues or groups of residues or even entire CDRs do not significantly impact antigen binding. The methods used herein may utilize CDRs defined according to any of these systems, although certain embodiments use Kabat-or Chothia-defined CDRs. “Communicated” or “communicating” as used herein refers to the conveying, transmiting and / or reporting of an item of information. In some embodiments, the information that is communicated is an item of information obtained by performing an assay, such as, the amount or presence of a biomarker(e.g., cMyc)in a sample (e.g., a result). The information obtained by performing an assay can be communicated by a computer, in a document and / or spreadsheet, on a mobile device (e.g., a smart phone), on a website,in an e-mail, or any combination thereof. In some other embodiments, information is communicated on or from an instrument or device. In other embodiments, the information is communicated by being displayed, such as on an instrument or device. As used herein, the term “control” refers to a reference standard for an analyte such as is known or accepted in the art, or determined empiricaly using acceptable means such as are commonly employed. A “reference standard” is a standardized substance which is used as a measurement base for a similar substance. For example, there are documented reference standards published in the U.S. Pharmacopeial Convention (USP–NF), Food Chemicals Codex, and Dietary Supplements Compendium (al of which are available at htp: / www.usp.org), and Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 other wel-known sources. Methods for standardizing references are described in the literature. Also wel-known are means for quantifying the amounts of analyte present by use of a calibration curve for analyte or by comparison to an alternate reference standard. A standard curve can be generated using serial dilutions or solutions of known concentrations of analyte, by mass spectroscopy, gravimetric methods, and by other techniques known in the art. Alternate reference standards that have been described in the literature include standard addition (also known as the method of standard addition), or digital polymerase chain reaction. “Derivative” of an antibody as used herein may refer to an antibody having one or more modifications to its amino acid sequence when compared to a genuine or parent antibody exhibita modified domain structure. The derivative may stil be able to adopt the typical domain configuration found in native antibodies, as wel as an amino acid sequence, which is able to bind to targets (antigens) with specificity. Typical examples of antibody derivatives are antibodies coupled to other polypeptides, rearanged antibody domains, or fragments of antibodies. The derivative may also comprise at least one further compound, e.g.,a protein domain, said protein domain being linked by covalent or non-covalent bonds. The linkage can be based on genetic fusion according to the methods known in the art. The additional domain present in the fusion protein comprising the antibody may preferably be linked by a flexible linker, advantageously a peptide linker, wherein said peptide linker comprises plural, hydrophilic, peptide-bonded amino acids of a length suficient to span the distance between the C-terminal end of the further protein domain and the N-terminal end of the antibody or vice versa. The antibody may be linked to an effector molecule having a conformation suitable for biological activity or selective binding to a solid support, a biologicaly active substance (e.g., a cytokine or growth hormone), a chemical agent, a peptide, a protein, or a drug, for example. “Dual-specific antibody” is used herein to refer to a ful-length antibody that can bind two diferent antigens (or epitopes) in each of its two binding arms (a pair of HC / LC) (see PCT publication WO 02 / 02773). Accordingly, a dual-specific binding proteinhas two identical antigen binding arms, with identical specificity and identical CDR sequences, and is bivalent for each antigen to which it binds. “Dual variable domain” is used herein to refer to two or more antigen binding sites on a binding protein, which may be divalent (two antigen binding sites), tetravalent (four antigen binding sites), or multivalent binding proteins. DVDs may be monospecific, i.e., capable of Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 binding one antigen (or one specific epitope), or multispecific, i.e., capable of binding two or more antigens (i.e.,two or more epitopes of the same target antigen molecule or two or more epitopes of diferent target antigens). A prefered DVD binding protein comprises two heavy chain DVD polypeptides and two light chain DVD polypeptides and is referred to as a “DVD immunoglobulin” or “DVD-Ig.” Such a DVD-Ig binding protein is thus tetrameric and reminiscent of an IgG molecule butprovides more antigen binding sites than an IgG molecule. Thus, each half of a tetrameric DVD-Ig molecule is reminiscent of one half of an IgG molecule and comprises a heavy chain DVD polypeptide and a light chain DVD polypeptide, but unlike a pair of heavy and light chains ofan IgG molecule that provides a single antigen binding domain, a pair of heavy and light chains of a DVD-Ig provide two or more antigen binding sites. Each antigen binding site of a DVD-Ig binding protein may be derived from a donor ("parental") monoclonal antibody and thus comprises a heavy chain variable domain (VH) and a light chain variable domain (VL) with a total of six CDRs involved in antigen binding per antigen binding site. Accordingly, a DVD-Ig binding protein that binds two diferent epitopes (i.e.,two diferent epitopes of two diferent antigen molecules or two diferent epitopes of the same antigen molecule) comprises an antigen binding site derived from a first parental monoclonal antibody and an antigen binding site of a second parental monoclonal antibody. A description of the design, expression, and characterization of DVD-Ig binding molecules is provided in PCT Publication No. WO 2007 / 024715, U.S. Patent No.7,612,181, and Wu et al., Nature Biotech., 25: 1290-1297 (2007). A prefered example of such DVD-Ig molecules comprises a heavy chain that comprises the structural formula VD1-(X1)n-VD2-C- (X2)n, wherein VD1 is a first heavy chain variable domain, VD2 is a second heavy chain variable domain, C is a heavy chain constant domain, X1 is a linker with the proviso that it is not CH1, X2 is an Fc region, and n is 0 or 1, but preferably 1; and a light chain that comprises the structural formula VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first light chain variable domain, VD2 is a second light chain variable domain,C is a light chain constant domain, X1 is a linker with the proviso that it is not CH1, and X2 does not comprise an Fc region; and n is 0 or 1, but preferably 1. Such a DVD-Ig may comprise two such heavy chains and two such light chains, wherein each chain comprises variable domains linked in tandem without an intervening constant region between variable regions, wherein a heavy chain and a light chain associate to form tandem functional antigen binding sites, and a pair of heavy and light chains may associate with Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 another pair of heavy and light chains to form a tetrameric binding protein with four functional antigen binding sites. In another example, a DVD-Ig molecule may comprise heavy and light chains that each comprise three variable domains (VD1, VD2, VD3) linked in tandem without an intervening constant region between variable domains, wherein a pair of heavy and light chains may associate to form three antigen binding sites, and wherein a pair of heavy and light chains may associate with another pair of heavyand light chains to form a tetrameric binding protein with six antigen binding sites. In a preferred embodiment, a DVD-Ig binding protein not only binds the same target molecules bound by its parental monoclonal antibodies, but also possesses one or more desirable properties of one or more of its parental monoclonal antibodies. Preferably,such an additional property is an antibody parameter of one or more of the parental monoclonal antibodies. Antibody parameters that may be contributed to a DVD-Ig binding protein from one or more of its parental monoclonal antibodies include, but are notlimited to, antigen specificity, antigen afinity, potency, biological function, epitope recognition, protein stability, protein solubility, production eficiency, immunogenicity, pharmacokinetics, bioavailability, tissue cross reactivity, and orthologousantigen binding. “Epitope,” or “epitopes,” or “epitopes of interest” refer to a site(s) on any molecule that is recognized and can bind to a complementary site(s) on its specific binding partner. The molecule and specific binding partner are part of a specific binding pair. For example, an epitope can be on a polypeptide, a protein, a hapten, a carbohydrate antigen (such as, but not limited to, glycolipids, glycoproteins or lipopolysaccharides), or a polysaccharide. Its specific binding partner can be, but is not limitedto, an antibody. “Fragment antigen-binding fragment” or “Fab fragment” as used herein refers toa fragment of an antibody that binds to antigens and that contains one antigen-binding site, one complete light chain, and part of one heavy chain. Fab is a monovalent fragment consisting of the VL, VH, CL and CH1 domains. Fab is composed of one constant and one variable domain of each of the heavy and the light chain. The variable domain contains the paratope (the antigen- binding site), comprising a set of complementarity determining regions, at the amino terminal end of the monomer. Each arm of the Y thus binds an epitope on the antigen. Fab fragments can be generated such as has been described in the art, e.g., using the enzyme papain, which can be Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 used to cleave an immunoglobulin monomer into two Fab fragments and an Fc fragment, or can be produced by recombinant means. “F(ab')2fragment” as used herein refers to antibodies generated by pepsin digestion of whole IgG antibodies to remove most of the Fc region while leaving intact some of the hinge region. F(ab')2fragments have two antigen-binding F(ab) portions linked together by disulfide bonds, and therefore are divalent with a molecular weight of about 110 kDa. Divalent antibody fragments (F(ab')2fragments) are smaler than whole IgG molecules and enable a beter penetration into tissue thus facilitating beter antigen recognition in immunohistochemistry. The use of F(ab')2fragments also avoidunspecific binding to Fc receptor on live cels or to Protein A / G. F(ab')2fragments can both bind and precipitate antigens. “Framework” (FR) or “Framework sequence” as used herein may mean the remaining sequences of a variable region minus the CDRs. Because the exact definition of a CDR sequence can be determined by diferent systems (for example, see above), the meaning of a framework sequence is subject to corespondingly diferent interpretations. The six CDRs (CDR-L1, -L2, and -L3 of light chain and CDR-H1, -H2, and -H3 of heavy chain) also divide the framework regions on the light chain and the heavy chain into four sub-regions (FR1, FR2, FR3, and FR4) on each chain, in which CDR1 is positioned between FR1 and FR2, CDR2 between FR2 and FR3, and CDR3 between FR3 and FR4. Without specifying the particular sub-regions as FR1, FR2, FR3, or FR4, a framework region, as refered toby others, represents the combined FRs within the variable region of a single, naturaly occuring immunoglobulin chain. As used herein, a FR represents one of the four sub-regions, and FRs represents two or more of the four sub-regions constituting aframework region. Human heavy chain and light chain FR sequences are known in the art that can be used as heavy chain and light chain "acceptor" framework sequences (or simply, "acceptor" sequences) to humanize a non-human antibody using techniques known in the art. In one embodiment, human heavy chain and light chain acceptor sequences are selected from the framework sequences listed in publicly available databases such as V-base (hypertext transfer protocol: / vbase.mrc-cpe.cam.ac.uk / ) or in the international ImMunoGeneTics® (IMGT®) information system (hypertext transfer protocol: / imgt.cines.fr / texts / IMGTrepertoire / LocusGenes / ). Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 “Functional antigen binding site” as used herein may mean a site on a binding protein (e.g.,an antibody) that is capable of binding a target antigen. The antigen binding afinity of the antigen binding site may not be as strong as the parent binding protein, e.g.,parent antibody, from which the antigen binding site is derived, but the ability to bind antigen must be measurable using any one of a variety of methods known for evaluating protein, e.g.,antibody, binding to an antigen. Moreover, the antigenbinding afinity of each of the antigen binding sites of a multivalent protein, e.g.,multivalent antibody, herein need not be quantitatively the same. “Humanized antibody” is used herein to describe an antibody that comprises heavy and light chain variable region sequences from a non-human species (e.g.,a mouse) but in which at least a portion of the VH and / or VL sequence has been altered to be more “human-like,” i.e., more similar to human germline variable sequences. A "humanized antibody" is an antibody or a variant, derivative, analog, or fragment thereof, which immunospecificaly binds to an antigen of interest and which comprises a framework (FR) region having substantialy the amino acid sequence of a human antibody and a complementary determining region (CDR) having substantialy the amino acid sequence of a non-human antibody. As used herein, the term "substantialy" in the context of a CDR refers to a CDR having an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% identical to the amino acid sequence of a non-human antibody CDR. A humanized antibody comprises substantialy al of at leastone, and typicaly two, variable domains (Fab, Fab', F(ab')2, FabC, Fv) in which al or substantialy al of the CDR regions corespond to those of a non-human immunoglobulin (i.e., donor antibody) and al or substantialy al of the framework regions are those of a human immunoglobulin consensus sequence. In an embodiment, a humanized antibody also comprises at least a portion of an immunoglobulin constant region (Fc), typicaly that of a human immunoglobulin. In some embodiments, a humanized antibody contains the light chain as wel as at least the variable domain of a heavy chain. The antibody also may include the CH1, hinge, CH2, CH3, and CH4 regions of the heavy chain. In some embodiments, a humanized antibody only contains a humanized light chain.In some embodiments, a humanized antibody only contains a humanized heavy chain. In specific embodiments, a humanized antibody only contains a humanized variable domain of a light chain and / or humanized heavy chain. A humanized antibody can be selected from any class of immunoglobulins, including IgM, IgG, IgD, IgA, and IgE, and any isotype, including without limitation IgG1, IgG2, IgG3, Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 and IgG4. A humanized antibody may comprise sequences from more than one class or isotype, and particular constant domains may be selected to optimize desired effector functions using techniques wel-known in the art. The framework regions and CDRs of a humanized antibody need not correspond precisely to the parental sequences, e.g.,the donor antibody CDR or the consensus framework may be mutagenized by substitution, insertion, and / or deletion of at least one amino acid residue so that the CDR or framework residue at that site does not corespond to either the donor antibody or the consensus framework. In a preferred embodiment, such mutations, however, wil not be extensive. Usualy, at least 80%, preferably at least85%, more preferably at least 90%, and most preferably at least 95% of the humanized antibody residues wil corespond to those of the parental FR and CDR sequences. As used herein, the term "consensus framework" refers to the framework region in the consensus immunoglobulin sequence. As used herein, the term "consensus immunoglobulin sequence" refers to the sequence formed from the most frequently occuring amino acids (or nucleotides) in a family of related immunoglobulin sequences (see, e.g.,Winnaker, From Genes to Clones(Verlagsgeselschaft, Weinheim, 1987). A "consensus immunoglobulin sequence" may thus comprise a "consensus framework region(s)" and / or a "consensus CDR(s)". In a family of immunoglobulins, each position in the consensus sequence is occupied by the amino acid occuring most frequently at that position in the family. If two amino acids occur equaly frequently, either can be included in the consensus sequence. “1A4” or “monoclonal antibody 1A4” as used interchangeably herein, refers to the monoclonal antibody described in U.S. Patent No.10,017,745, the contents of which are herein incorporated by reference. Monoclonal antibody 1A4specificaly binds to an epitope having the amino acid sequence of: AAELGESAPSPK(SEQ ID NO:2). In some embodiments, monoclonal antibody 1A4can be produced by hybridoma cel line deposited with the DSMZ and assigned Accession Number DSM ACC3224. In other embodiments, monoclonal antibody 1A4can be produced by recombinant means using recombinant DNA technology (e.g., the antibody is generated in vitrousing synthetic genes introduced into mammalian cel lines rather than through hybridoma cel culture or animal immunization), which is wel known in the art.In stil other embodiments, chimeric, CDR-grafted, humanized,fuly human, andafinity matured versions of 1A4can be produced and used in the methods described herein using routine techniques known in the art. Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 “Identical” or “identity,” as used herein in the context of two or more polypeptide or polynucleotide sequences, can mean that the sequences have a specified percentage of residues that are the same over a specified region. The percentage can be calculated by optimaly aligning the two sequences, comparing the two sequences over the specified region, determining the number of positions at which the identical residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the specified region, and multiplying the result by 100 to yield the percentage of sequence identity. In cases where the two sequences are of diferent lengths or the alignment produces one or more staggered ends and the specified region of comparison includes only a single sequence, the residues of the single sequence are included in the denominator but not the numerator of the calculation. “Isolated polynucleotide” as used herein may mean a polynucleotide (e.g., of genomic, cDNA, or synthetic origin, or a combination thereof) that, by virtue of its origin, the isolated polynucleotide is not associated with al or a portion of a polynucleotide with which the “isolated polynucleotide” is found in nature; is operably linked to a polynucleotide that it is not linked to in nature; or does not occur in nature as part of a larger sequence. “Label” and “detectable label” as used herein refer to a moiety atached to an antibody or an analyte to render the reaction between the antibody and the analyte detectable, and the antibody or analyte so labeled is refered to as “detectably labeled.” A label can produce a signal that is detectable by visual or instrumental means. Various labels include signal-producing substances, such as chromagens, fluorescent compounds, chemiluminescent compounds, radioactive compounds, and the like. Representative examples of labels include moieties that produce light, e.g., acridinium compounds, and moieties that produce fluorescence, e.g., fluorescein. Other labels are described herein. In this regard, the moiety itselfmay not be detectable but may become detectable upon reaction with yet another moiety. Use of the term “detectably labeled” is intended to encompass such labeling. Any suitable detectable label as is known in the art can be used. For example, the detectable label can be a radioactive label (such as 3H, 14C, 32P, 33P, 35S, 90Y, 99Tc, 111In, 125I, 131I, 177Lu, 166Ho, and 153Sm), an enzymatic label (such as horseradishperoxidase, alkaline peroxidase, glucose 6-phosphate dehydrogenase, and the like), a chemiluminescent label (such as acridinium esters, thioesters, or sulfonamides; luminol, isoluminol, phenanthridinium Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 esters, and the like), a fluorescent label (such as fluorescein (e.g., 5-fluorescein, 6- carboxyfluorescein, 3’6-carboxyfluorescein, 5(6)-carboxyfluorescein, 6-hexachloro-fluorescein, 6-tetrachlorofluorescein, fluorescein isothiocyanate, and the like)), rhodamine, phycobiliproteins, R-phycoerythrin, quantum dots (e.g., zinc sulfide-capped cadmium selenide), a thermometric label, or an immuno-polymerase chain reaction label. An introduction to labels, labeling procedures and detection of labels is found in Polak and Van Noorden, Introduction to Immunocytochemistry, 2nd ed., Springer Verlag, N.Y. (1997), and in Haugland, Handbook of Fluorescent Probes and Research Chemicals(1996), which is a combined handbook and catalogue published by Molecular Probes, Inc., Eugene, Oregon. A fluorescent label can be used in FPIA (see, e.g., U.S. Patent Nos.5,593,896, 5,573,904, 5,496,925, 5,359,093, and 5,352,803, which are hereby incorporated by reference in their entireties). An acridinium compound can be used as a detectable label in a homogeneous chemiluminescent assay (see, e.g., Adamczyk et al., Bioorg. Med. Chem. Let.16: 1324-1328 (2006); Adamczyk et al.,Bioorg. Med. Chem. Let.4: 2313-2317 (2004); Adamczyk et al., Biorg. Med. Chem. Let.14: 3917-3921 (2004); and Adamczyk et al.,Org. Let.5: 3779-3782 (2003). In one embodiment, the acridinium compound is an acridinium-9-carboxamide. Methods for preparing acridinium 9-carboxamides are described in Matingly, J. Biolumin. Chemilumin.6: 107-114 (1991); Adamczyk et al.,J. Org. Chem.63: 5636-5639 (1998); Adamczyk et al., Tetrahedron 55: 10899-10914 (1999); Adamczyk et al., Org. Let.1: 779-781 (1999); Adamczyk et al.,Bioconjugate Chem.11: 714-724 (2000); Matingly et al.,In Luminescence Biotechnology: Instruments and Applications; Dyke, K. V. Ed.; CRC Press: Boca Raton, pp.77–105 (2002); Adamczyk et al.,Org. Let.5: 3779-3782 (2003); and U.S. Patent Nos.5,468,646, 5,543,524 and 5,783,699 (each of which is incorporated herein by reference in its entirety for its teachings regarding same). Another example of an acridinium compound is an acridinium-9-carboxylate aryl ester. An example of an acridinium-9-carboxylate aryl ester of formula I is 10-methyl-9- (phenoxycarbonyl)acridinium fluorosulfonate(available from Cayman Chemical, Ann Arbor, MI). Methods for preparing acridinium 9-carboxylate aryl esters are described in McCapra et al., Photochem. Photobiol.,4: 1111-21 (1965); Razavi et al.,Luminescence15: 245-249 (2000); Razavi et al., Luminescence15: 239-244 (2000); and U.S. Patent No.5,241,070 (each of which is incorporated herein by reference in its entirety for its teachings regarding same). Such Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 acridinium-9-carboxylate aryl esters are eficient chemiluminescent indicators for hydrogen peroxide produced in the oxidation of an analyte by at least one oxidase in terms of the intensity of the signal and / or the rapidity of the signal. The course of the chemiluminescent emission for the acridinium-9-carboxylate aryl ester is completed rapidly, i.e., in under 1 second, while the acridinium-9-carboxamide chemiluminescent emission extends over 2 seconds. Acridinium-9- carboxylate aryl ester, however, loses its chemiluminescent properties in the presence of protein. Therefore, its use requires the absence of protein during signal generation and detection. Methods for separating or removing proteins in the sample are wel-known to those skiled in the art and include, but are not limited to, ultrafiltration, extraction, precipitation, dialysis, chromatography, and / or digestion (see, e.g.,Wels, High Throughput Bioanalytical Sample Preparation. Methods and Automation Strategies, Elsevier (2003)). The amount of protein removed or separated from the test sample can be about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95%. Further details regarding acridinium-9-carboxylate aryl ester and its use are set forth in U.S. Patent App. No.11 / 697,835, filed April 9, 2007. Acridinium-9-carboxylate aryl esters can be dissolved in any suitable solvent, such as degassed anhydrous N,N-dimethylformamide (DMF) or aqueous sodium cholate. As used herein, the term “microparticle(s)” refers tosmal particles with dimensions typicaly ranging from 0.1 to 100 micrometers (μm).These particles can be solid or coloidal in nature. Microparticles can include,but are not limited to, polymeric microparticles, liposomes, microspheres, nanoparticles, magnetic microparticles, non-magnetic microparticles, protein microparticles, biodegradable microparticles, ceramic microparticles, holow microparticles, Janus particles, nanospheres, microcapsules, and nanocapsules. In some cases, microparticle can include one or more of the folowing: apoly(lactide-co-glycolide), aliphatic polyesters including, but not limited to, poly-glycolic acid andpoly-lactic acid, hyaluronic acid, modified polysaccharides, chitosan, celulose, dextran,polyurethanes, polyacrylic acids, pseudo- poly(amino acids),polyhydroxybutyrate-relatedcopolymers, polyanhydrides, polymethylmethacrylate, poly(ethylene oxide), lecithin and phospholipids –in any combination thereof. “Non-point-of-care device” refers to a device that is not a point-of-care device or a single use device. A non-point-of-care device refers to any device that does not meet any of the Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 above limitations of a point-of-care or a single use deviceas defined herein. In some embodiments, the non-point-of-care device may be a relatively large instrument, such as a tabletop instrument. Accordingly, in some embodiments the non-point-of-care device is not a handheld instrument. In some embodiments, the non-point-of-care device is capable of performing an assay on more than one clinical sample simultaneously. Suitable non-point-of-care devices include, for example, the Architect or Alinity platforms produced by Abbot Core Laboratories. “Point-of-care device” refers to a device used to provide medical diagnostic testing at or near the point-of-care (namely, outside of a laboratory), at the time and place of patient care (such as in a hospital, physician’s ofice, urgent or other medical care facility, a patient’s home, a nursing home and / or a long-term care and / or hospice facility). Examples of point-of-care devices include those produced by Abbot Laboratories (Abbot Park, IL) (e.g.,i-STAT and i- STAT Alinity, Universal Biosensors (Rowvile, Australia) (see US 2006 / 0134713), Axis-Shield PoC AS (Oslo, Norway) and Clinical Lab Products (Los Angeles, USA). In some embodiments, the point-of-care device is a single-use device. The term “single-use device” or “single-use instrument” refers to a clinical diagnostic instrument that processes and performs a clinical diagnostic assay on a unit use basis (such as, for example, a single-use cartridge) for a single patient sample. A point-of-care instrument does not perform an assay on more than oneclinical sample simultaneously. However, the point-of-care instrument may have the capability to measure more than one parameter (e.g., more than one analyte) in an individual clinical sample per unit use basis. “Quality control reagents” in the context of immunoassays and kits described herein, include, but are not limited to, calibrators, controls, and sensitivity panels. A “calibrator” or “standard” typicaly is used (e.g., one or more, such as a plurality) inorder to establish calibration (standard) curves for interpolation of the concentration of an analyte, such as an antibody or an analyte. Alternatively, a single calibrator, which is near a reference level or control level (e.g., “low”, “medium”, or “high” levels), can be used. Multiple calibrators (i.e., more than one calibrator or a varying amount of calibrator(s)) can be used in conjunction to comprise a “sensitivity panel.” “Recombinant antibody” and “recombinant antibodies” refer to antibodies prepared by one or more steps, including cloning nucleic acid sequences encoding al or a part of one or more Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 monoclonal antibodies into an appropriate expression vector by recombinant techniques and subsequently expressing the antibody in an appropriate host cel. The terms include, but are not limited to, recombinantly produced monoclonal antibodies, chimeric antibodies, humanized antibodies (fuly or partialy humanized), multi-specific or multi-valent structures formed from antibody fragments, bifunctional antibodies, heteroconjugate Abs, DVD-Ig®s, and other antibodies as described in (i) herein. (Dual-variable domain immunoglobulins and methods for making them are described in Wu, C., et al., Nature Biotechnology, 25:1290-1297 (2007). The term “bifunctional antibody,” as used herein, refers to an antibody that comprises a first arm having a specificity for one antigenic site and a second arm having a specificity for a diferent antigenic site, i.e., the bifunctional antibodies have a dual specificity. “Reference level” as used herein refers to an assay cutof value that is used to assess diagnostic, prognostic, or therapeutic eficacy and that has been linked or is associated herein with various clinical parameters (e.g., presence of disease, stage of disease, severity of disease, progression, non-progression, or improvement of disease, etc.). This disclosure provides exemplary reference levels. However, it is wel-known that reference levels may vary depending on the nature of the immunoassay (e.g., antibodies employed, reaction conditions, sample purity, etc.) and that assays can be compared and standardized. It further is wel within the ordinary skil of one in the art to adapt the disclosure herein for other immunoassays to obtain immunoassay-specific reference levels for those other immunoassays based on the description provided by this disclosure. Whereas the precise value of the reference level may vary between assays, the findings as described herein should be generaly applicable and capable of being extrapolated to other assays. In certain embodiments described herein, the reference level is described as being determined by any assay having a certain specificity and sensitivity. “Sample,” “test sample,” “specimen,” “sample from a subject,” “biological sample,” and “patient sample” as used interchangeably herein may be a sample of blood, such as whole blood (including for example, capilary blood, venous blood, dried blood spot, etc.), serum or plasma, or tissue, saliva, urine, , amniotic fluid, an oropharyngeal specimen, a nasopharyngeal specimens, lower respiratory specimens such as, but not limited to, sputum, endotracheal aspirate or bronchoalveolar lavage, cerebrospinal fluid, placental cels or tissue, endothelial cels, leukocytes, or monocytes. The sample can be used directly as obtained from a patient or can be pre-treated, such as by filtration, distilation, extraction, concentration, centrifugation, Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 inactivation of interfering components, addition of reagents, and the like, to modify the character of the sample in some manner as discussed herein or otherwise as is known in the art. Additionaly, the sample can be a nasopharyngeal or oropharyngeal sample obtained using one or more swabs that, once obtained, is placed in a sterile tube containing a virus transport media (VTM) or universal transport media (UTM), and retained therein or transfered to another media for testing. A variety of cel types, tissue, or bodily fluid may be utilized to obtain a sample. Such cel types, tissues, and fluid may include sections of tissues such as biopsy and autopsy samples, oropharyngeal specimens, nasopharyngeal specimens, frozen sectionstaken for histologic purposes, blood (such as whole blood, dried blood spots, etc.), plasma, serum, saliva, red blood cels, platelets, interstitial fluid, cerebral spinal fluid, etc. Cel types and tissues may also include lymph fluid, cerebrospinal fluid, or any fluid colected by aspiration. A tissue or cel type may be provided by removing a sample of cels from a human and a non-human animal but can also be accomplished by using previously isolated cels (e.g., isolated by another person, at another time, and / or for another purpose). Archival tissues, such as those having treatment or outcome history, may also be used. Protein or nucleotide isolation and / or purification may not be necessary. In some embodiments, the sample is a blood sample (e.g., a whole blood sample, a serum sample, or a plasma sample). In some embodiments, the sample is a whole blood sample. In some embodiments, the sample is a capilary blood sample. In some embodiments, the sample is a dried blood spot. In some embodiments, the sample is a serum sample. In yet other embodiments, the sample is a plasma sample. In some embodiments, the sample is an oropharyngeal specimen. In other embodiments, the sample is a nasopharyngeal specimen. In other embodiments, the sample is sputum. In other embodiments, the sample is endotracheal aspirate. In stil yet other embodiments, the sample is bronchoalveolar lavage. In stil yet other embodiments, the sample is a saliva sample. “Sensitivity” of an assay as used herein refers to the proportion of subjects for whom the outcome is positive that are corectly identified as positive (e.g., correctly identifying those subjects with a disease or medical condition for which they are being tested). For example, this might include corectly identifying subjects as having a cardiac injuryas distinct from those who do not have a cardiac injury. Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 “Specificity” of an assay as used herein refers to the proportion of subjects for whom the outcome is negative that are corectly identified as negative (e.g., correctly identifying those subjects who do not have a disease or medical condition for which they are being tested). For example, this might include corectly identifying subjects not having acardiac injury as distinct from those who do have a cardiac injury. “Solid phase” or “solid support” as used interchangeably herein, refers to any material that can be used to atach and / or atract and immobilize (1) one or more capture reagents or capture specific binding partners, or (2) one or more detection reagents ordetection specific binding partners. The solid phase can be chosen for its intrinsic ability to atract and immobilize a capture reagent. Alternatively, the solid phase can have afixed thereto a linking agent that has the ability to atract and immobilize the (1) capture reagent or capture specific binding partner, or (2) detection reagent or detection specific binding partner. For example, the linking agent can include a charged substance that is oppositely charged with respect to the capture reagent (e.g., capture specific binding partner) or detection reagent (e.g.,detection specific binding partner) itself or to a charged substance conjugated to the (1) capture reagent or capture specific binding partner or (2) detection reagent or detection specific binding partner. In general, the linking agent can be any binding partner (preferably specific) that is immobilized on (atached to) the solid phase and that has the ability to immobilize the (1) capture reagent or capture specific binding partner, or (2) detection reagent or detection specific binding partner through a binding reaction. The linking agent enables the indirect binding of the capture reagent to a solid phase material before the performance of the assay or during the performance of the assay. For example, the solid phase can be plastic, derivatized plastic, magnetic, or non-magnetic metal, glass or silicon, including, for example, a test tube, microtiter wel, sheet, bead, microparticle, chip, and other configurations known to those of ordinary skil in the art. In some embodiments, the solid phase is polystyrene or derivatized polystyrene.In some embodiments, a solid support is a bead or a particle, such as a microparticle. “Specific binding” or “specificaly binding” as used herein may refer to the interaction of an antibody, a protein, or a peptide with a second chemical species, wherein the interaction is dependent upon the presence of a particular structure (e.g.,an antigenic determinant or epitope) on the chemical species; for example, an antibody recognizes and binds to a specific protein structure rather than to proteins generaly. If an antibody is specific for epitope “A”, the Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 presence of a molecule containing epitope A (or free, unlabeled A), in a reaction containing labeled “A” and the antibody, wil reduce the amount of labeled A bound to the antibody. “Specific binding partner” is a member of a specific binding pair. A specific binding pair comprises two diferent molecules, which specificaly bind to each other through chemical or physical means. Therefore, in addition to antigen and antibody specific binding pairs of common immunoassays, other specific binding pairs can include biotin and avidin (or streptavidin), carbohydrates and lectins, complementary nucleotide sequences, effector and receptor molecules, cofactors and enzymes, enzymes and enzyme inhibitors, and the like. Furthermore, specific binding pairs can include members that are analogs of the original specific binding members, for example, an analyte-analog. Immunoreactive specific binding members include antigens, antigen fragments, and antibodies, including monoclonal and polyclonal antibodies as wel as complexes and fragments thereof, whether isolated or recombinantly produced, and aptamers. “Subject” and “patient” as used herein interchangeably refers to any vertebrate, including, but not limited to, a mammal (e.g.,cow, pig, camel, lama, horse, goat, rabbit, sheep, hamsters, guinea pig, cat, dog, rat, and mouse, a non-human primate (for example, a monkey, such as a cynomolgus or rhesus monkey, chimpanzee, etc.) and a human). In some embodiments, the subject may bea human or a non-human. In some embodiments, the subject is a human. The subject or patient may be undergoing other forms of treatment. “Treat,” “treating” or “treatment” are each used interchangeably herein to describe reversing, aleviating, or inhibiting the progress of a disease or one or more symptoms of such disease, to which such term applies. In some aspects, a treatment may be either performed in an acute or chronic way. Depending on the condition of the subject, the term also refers to preventing a disease and includes preventing the onset of a disease, or preventing the symptoms associated with a disease. "Preventing" also refers to preventing the recurence of a disease or of one or more symptoms associated with such disease. "Treatment" and "therapeuticaly," refer to the act of treating, as "treating" is defined above. In some aspects, the prevention or treatment of a disease can be done prior to afliction or injury, such as, for example, to reduce the severity of a disease or symptoms associated with a disease. Such prevention or reduction can include (a) administration of one or more pharmaceutical composition and / or one or more nutritional compositions to a subject; (b) the use of one or more surgical interventions (e.g., angioplasty, Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 inserting a stent, atherectomy, bypass surgery or any combinations thereof); or (c) any combinations of (a) and (b). “Variant” is used herein to describe a peptide or polypeptide that difers in amino acid sequence by the insertion, deletion, or conservative substitution of amino acids, but retain at least one biological activity. Representative examples of “biological activity” include the ability to be bound by a specific antibody or to promote an immune response. Variant is also used herein to describe a protein with an amino acid sequence that is substantialy identical to a referenced protein with an amino acid sequence that retains at least one biological activity. A conservative substitution of an amino acid, i.e.,replacing an amino acid with a diferent amino acid of similar properties (e.g.,hydrophilicity, degree, and distribution of charged regions) is recognized in the art as typicaly involving a minor change. These minor changes can be identified, in part, by considering the hydropathic index of amino acids, as understood in the art. Kyte et al.,J. Mol. Biol.157:105-132 (1982). The hydropathic index of an amino acid is based on a consideration of its hydrophobicity and charge. It is known in the art that amino acids of similar hydropathic indexes can be substituted and stil retain protein function. In one embodiment, amino acids having hydropathic indexes of ±2 are substituted. The hydrophilicity of amino acids can also be used to reveal substitutions that would result in proteins retaining biological function. A consideration of the hydrophilicity of amino acids in the context of a peptide permits calculation of the greatest local average hydrophilicity of that peptide, a useful measure that has been reported to corelate wel with antigenicity and immunogenicity. U.S. Patent No.4,554,101, incorporated fuly herein by reference. Substitution of amino acids having similar hydrophilicity values can result in peptides retaining biological activity, for example immunogenicity, as is understood in the art. Substitutions may be performed with amino acids having hydrophilicity values within ±2 of each other. Both the hydrophobicity index and the hydrophilicity value of amino acids are influenced by the particular side chain of that amino acid. Consistent with that observation, aminoacid substitutions that are compatible with biological function are understood to depend on the relative similarity of the amino acids, and particularly the side chains of those amino acids, as revealed by the hydrophobicity, hydrophilicity, charge, size, and other properties. 2. Assays and Methods of Measuring a Level of a Cardiac Myosin Binding Protein C in Biological Samples obtained from a Subject Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 In some embodiments, provided herein are assays and methods for measuring an amount, level,or concentrationof a cardiac myosin binding protein C (“cMyC”) in at least one biological sample obtained from a subject. In some embodiments, the subject is sufering from or suspected of having sufered from a cardiac injury. In yet another embodiment, the subject is sufering from or suspected of sufering from myocardial infarction. In some embodiments, the myocardial infarction is an acute myocardial infarction. In other embodiments, the myocardial infarction is a chronic myocardial infarction. In stil other embodiments, the sufering from or suspected of suffering from a reinfarction. In some embodiments, the methods described herein can be used to diagnose or detect a cardiac injury in a subject. In some embodiments, the methods described herein can be used to diagnose or detect a myocardial infarction in a subject. In yet other embodiments, the methods described herein can be used to diagnose or detect an acute myocardial infarction in a subject.In stil yet other embodiments, the methods described herein can be used to diagnose or detect a chronic myocardial infarction in a subject. In stil yet other embodiments, the methods described herein can be used to diagnose or detect a reinfarction in a subject. In some embodiments, the method comprises obtaining or receiving a level of cMyC determined for a subject. As mentioned previously, the subject may be suffering from or suspected of having sufered from a cardiac injury such as a myocardial infarction or reinfarction. In some embodiments, the subject mayexhibitone or more symptoms of a myocardial infarctionor reinfarction, such as uncomfortable pressure, squeezing, fulness, stabbing pain, chest discomfort, discomfort in other areas of the upper body (suchas one or both arms, the back, neck, jaw, or stomach), shortness of breath with or without chest discomfort, shortnessof breath with or without chest discomfort, pounding heart or changes in heart rhythm, heartburn, nausea, vomiting, and abdominal pain, breaking out in a cold sweat or clammy skin or dizziness or lightheadness, body aches, sleep disturbance or anxiety. In some embodiments, the method comprises obtaining one or more biological samples from a subject sufering from or suspected of having sufered from a cardiac injury such as a myocardial infarction or reinfarction. In some embodiments, the biological sample is a whole blood sample. In other aspects, the biological sample is a plasma sample. In stil other aspects, the biological sample is a serum sample.Additionaly, in some embodiments, serial sampling from a subject is contemplated. Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 Oncethe at least one biological sample is obtained from the subject, at least one assay is performed. The assay involves contacting the at least one biological sample, either simultaneously or sequentialy, in any order with at least one first specific binding partner (e.g., “capture specific binding partner”) and at least one second specific binding partnerto forma first mixture comprising one or more first complexes comprising the at least one first specific binding partner-cMyC-at leastone second specific binding partner. In some embodiments, the at least one first specific binding partner and the at least one second specific binding partner specificaly bind to diferent epitopes on cMyC. In other words, the at least one first specific binding partner specificaly binds to a first epitope on cMyC and the at least one second specific binding partner specificaly binds to a second epitope on cMyC that is a diferent epitope than the epitope that the at least one first specific binding partner specificaly binds. In some embodiments, the at least one first specific binding partner is monoclonal antibody 3H8(e.g., “capture antibody”). In stil further embodiments, the at leastone first specific binding partner is immobilized on a solid support. In stil further embodiments, the solid support is a bead or particle, such as a microparticle(e.g., such a magnetic microparticle).In stil further embodiments, the at least one first specific binding partner is monoclonal antibody 3H8 which is immobilized on at least one solid support. In stil further embodiments, the at least one first specific binding partner is monoclonal antibody 3H8 which is immobilized on at least one microparticle.In stil further embodiments, the at least one first specific binding partner is monoclonal antibody 3H8 which is immobilized on at least one magnetic microparticle. In stil yet other embodiments, the at least one second specific binding partner comprises a detectable label (e.g., “detection specific binding partner”). In stil other embodiments, the at least second specific binding partner ismonoclonal antibody 1A4. In stil yet other embodiments, the at least second specific binding partner ismonoclonal antibody1A4 which furthercomprises a detectable label(e.g., “conjugate” or “detection antibody”). In stil yet other embodiments, the at least second specific binding partner is monoclonal antibody 1A4 which furthercomprises an acridinium label. In stil yet other embodiments, the at least one first specific binding partner is monoclonal antibody 3H8immobilized on a solid support and the at least one second specific binding partner is monoclonal antibody 1A4whichcomprises a detectable label. In stil yet other embodiments, the at least one first specific binding partner is monoclonal antibody 3H8 Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 immobilized on a microparticle and the at least one second specific binding partner is monoclonal antibody 1A4and comprises an acridinium label.In stil yet other embodiments, the at least one first specific binding partner is monoclonal antibody 3H8 immobilized on a magnetic microparticle and the at least one second specific binding partner is monoclonal antibody 1A4 and comprises an acridinium label. Once the at least one biological sample is contacted with the at least one first specific binding partner and the at least one second binding partner,the first mixture is alowed to incubate for about 12 to about 30 minutes,to alow one or more first complexes comprising the first specific binding partner-cMyC-second specific binding partner complexes to form.In some embodiments, the first mixture is alowed to incubate for about 20 minutes, about 19 minutes, about 18 minutes, about 17 minutes, about 16 minutes or about 15 minutes to alow one or more first complexes comprising the first specific binding partner-cMyC-second specific binding partner complexes to form.Once the incubation is completed, the at least one first specific binding partner-cMyC-second specific binding partner complexesare isolated from thefirst mixture. The complexes can be isolated using routine techniquesknownin the art. For example, if a magnetic microparticle is used with the at least one first specific binding partner,the microparticlescontaining the at least one first specific binding partner-cMyC-second specific binding partner complexescan be isolated or sequestered at the side of a reaction vessel using a magnetwhile al or portion of the first mixture is removed (e.g., in some embodiments, if the mixture is centrifuged, al or a portionof the supernatant can be removed). Once the at least one first specific binding partner-cMyC-second specific binding partner complexes are isolatedfrom the first mixture, the complex is washed with asolution to form a second mixture. The solution used for the washing(e.g., wash solution)can comprise water or at least one detergent. In some embodiments, the at least one detergentisa surfactant, such as an anionic surfactant, such as methyl ester sulfonate or a non-ionic surfactant such as tergitol,such as tergitol 15-S-40(Secondary alcohol ethoxylate with 41 ethylene oxide(EO) units).In some embodiments, the wash solution cancontain about a 20% concentration of at least one detergent. In other embodiments, theamount of tergitol that can be used in thewash solution is about 0.1% to about 2% v / v. After the one first specific binding partner-cMyC-second specific binding partner complexes is washed with the washsolution to form a second mixture, at least one bufer Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 solution can be added. The at least one bufer solution can contain 2-(N- morpholino)ethanesulfonic acid (MES), at least one salt, at least one protein blocker, at least one antimicrobial agent, and at least one detergent. The second mixture comprising the at least one first specific binding partner-cMyC- second specific binding partner complex is incubated for about 1 to about 5 minutes. In some embodiments, the second mixture comprising the at least one first specific binding partner- cMyC-second specific binding partner complex is incubated for about 4 minutes. Once the incubation is completed, the at least one first specific binding partner-cMyC-second specific binding partner complexesare isolated from the second mixture. These complexes can be isolated using routine techniques known in the art. For example, if a magnetic microparticle is used with the at least one first specific binding partner, the microparticles containing the at least one first specific binding partner-cMyC-second specific binding partner complexescan be isolated or sequestered at the side of a reaction vessel using a magnet while al or portion of the second mixture is removed (e.g., in some embodiments, if the mixture is centrifuged, al or part of the supernatant can be removed). Once the at least one first specific binding partner-cMyC-second specific binding partner complexes are isolated from the secondmixture, the complex is washed with a solution to form a second mixture. The solution used for the washing can comprise water or at least one detergent. In some embodiments, the at least one detergent is a surfactant, such as an anionic surfactant, such as methyl ester sulfonate or a non-ionic surfactant such as tergitol,such as tergitol 15-S-40 (Secondary alcohol ethoxylate with 41 ethylene oxide (EO) units). In some embodiments, the solution can contain about a 20% concentration of at least one detergent. In other embodiments, theamount of tergitol that can be used in the solution is about 0.1% to about 2% v / v. The final step of the method involves measuring the amount, level,or concentration of cMyC in the sample based upon the signal generated by the detectable label in the at least one first specific binding partner-cMyC-second specific binding partner complex using routine techniques known in the art. Once the amount or concentration of cMyC in the sample is determined, it can be communicated using routine techniques known in the art as described in more detail herein. Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 In some embodiments, the methods described hereinprovide an amount, level,or concentration of cMyC (e.g., generates a result) in a biological sample within about 1 hour from the start of the assay(e.g., the start of the assay is when the biological sample is first contacted with either at least first specific binding partner (e.g., monoclonal antibody 3H8)or second specific binding partner (e.g., monoclonal antibody 1A4).In other embodiments, In some embodiments, the methods described herein provide an amount, level, or concentration of cMyC (e.g., generates a result) in a biological sample within about 30 minutes from the start of the assay (e.g., the start of the assay is when the biological sample is first contacted with either at least first specific binding partner (e.g., monoclonal antibody 3H8) or second specific binding partner (e.g., monoclonal antibody 1A4). Areference level of cMyC can also be employed as a benchmark against which to assess results obtained upon assaying a test sample forcMyC. Generaly, in making such a comparison, the reference level ofcMyC, is obtained by running or conducting a particular assay a suficient number of times and under appropriate conditions such that a linkage or association of analyte presence, amount or concentration with a particular stage or endpoint ofcardiac injury. Typicaly, the reference level ofcMyC, is obtained with assays of reference subjects (or populations of subjects). cMyC measured can include fragments thereof, degradation products thereof, and / or enzymatic cleavage products thereof. In certain embodiments, the reference level may be corelated with control subjects (e.g., human subjects) that have not sustained a cardiac injury. In some embodiments, the method comprises performing at least one assay for a cMyC, and determining whether the subject’s levels of the cMyC are elevatedbased upon the results of the assay. In some embodiments, the levels are determined to be elevatedwhen compared to a reference level. In these embodiments, the subject can be determined to have a cardiac injury. In some embodiments, the levels are determined to not be elevatedwhen compared to a reference level.In these embodiments, the subject can be determined to not have a cardiac injury. In some embodiments, the method further includes treating a subject (e.g. a human subject) assessed as having a cardiac injury with a cardiac injury treatment, as described in Section 4. In some embodiments, the method further includes monitoring a subject (e.g., a human subject) assessed as having a cardiac injury, as describedin Section 4. Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 In some embodiments, the methodscomprise communicatingthe concentration or level of cMyC in a biological sample or adetermination of whether the level of cMyCin a biological sample obtained from a subject iselevatedornot elevatedwhen compared to a reference level. In some embodiments, the method comprises communicating the concentration or level of cMyC or a determination (e.g. the determination that subject’s levels of cMyC are elevatedorthe determination that the subject’s levels of cMyC are not elevatedwhen compared to a referencelevel) on or from at least one instrument. Suitable instruments are described herein, including point-of-care devicesand non-point-of care devicesthat may contain a user interface that communicatesby displaying the determination. As discussed in further detail in Section 3, in some embodiments, the instrument contains software to execute one or more tasks. In some embodiments, the instrument contains software to automaticaly determine the next appropriate step in a method as described herein. For example, the instrument maycontain software that determines whether levels of cMyC are elevatedor whether levels are not elevatedwhen compared to a reference level. The software may display this determination, such as on a graphical user interface. In some embodiments, the instrument stores software that instructs a processor to execute a given task. In some embodiments, the software stores machine readable instructions that instruct a processor to execute a given task. The machine-readable instructions may be one or more executable programs or portion(s) of an executable program for execution by a computer. The programs may be embodied in software stored on a non-transitory computer readable storage medium such as a CD-ROM, a floppy disk, a hard drive, a DVD, a Blu-ray disk, or a memory associated with the processors. Alternatively, the entire programs and / or parts thereof could alternatively be executed by a device other than the processors and / or embodied in firmware or dedicated hardware. Additionaly,or alternatively, processes may be implemented by one or more hardware circuits (e.g., discrete and / or integrated analog and / or digital circuitry, an FPGA, an ASIC, a comparator, an operational-amplifier (op-amp), a logic circuit, etc.) structured to perform the coresponding operation without executing software or firmware. The machine-readable instructions may be stored in one or more of a compressed format, an encrypted format, a fragmented format, a compiled format, an executable format, a packaged format, etc. Machine-readable instructions as described herein may be stored as data (e.g., portions of instructions, code, representations of code, etc.) that may be utilized to create, Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 manufacture, and / or produce machine executable instructions. For example, the machine- readable instructions may be fragmented and stored on one or more storage devices and / or computing devices (e.g., servers). The machine-readable instructions may require one or more of instalation, modification, adaptation, updating, combining, supplementing, configuring, decryption, decompression, unpacking, distribution, reassignment, compilation, etc. in order to make them directly readable, interpretable, and / or executable by a computing device and / or other machine. For example, the machine-readable instructions may be stored in multiple parts, which are individualy compressed, encrypted, and stored on separate computing devices, wherein the parts when decrypted, decompressed, and combined form a set of executable instructions that implement a program such as that described herein. In another example, the machine-readable instructions may be stored in a state in which they may be read by a computer, but require addition of a library (e.g., a dynamic link library (DLL), a software development kit (SDK), an application programming interface (API), etc. in order to execute the instructions on a particular computing device or other device. In another example, the machine-readable instructions may need to be configured (e.g., setings stored, data input, network addresses recorded, etc.)before the machine-readableinstructions and / or the corresponding program(s) can be executed in whole or in part. Thus, the disclosed machine-readable instructions and / or coresponding program(s) are intended to encompass such machine-readableinstructions and / or program(s) regardless of the particular format or state of the machine-readableinstructions and / or program(s) when stored or otherwise at rest or in transit. The machine-readable instructions described herein can be represented by any past, present, or future instruction language, scripting language, programming language, etc. For example, the machine-readable instructions may be represented using any of the folowing languages: C, C++, Java, C#, Perl, Python, JavaScript, HyperText Markup Language (HTML), Structured Query Language (SQL), Swift, etc. The machine-readable instructions may be stored on a non-transitory computer and / or machine readable medium such as a hard disk drive, a flash memory, a read-only memory, a compact disk, a digital versatile disk, a cache, a random-access memory and / or any other storage device or storage disk in which information is stored for any duration (e.g., for extended time periods, permanently, for brief instances, for temporarily bufering, and / or for caching of the Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 information). As used herein, the term non-transitory computer readable medium is expressly defined to include any type of computer readable storage device and / or storage disk and to exclude propagating signals and to exclude transmission media. Tests or assays competent to perform the claimed methods wil be employed, such as, for example, assays having various sensitivities and sensitivities as described herein. Moreover, the assays and methods described herein can be employed in an immunoassay orclinical chemistry format such as would be known by one of ordinary skil in the art. Moreover, the assays and methods described herein can also be performed using single molecule detection, lateral flow, or a point-of-care method. It is known in the art that the values (e.g., reference levels, cutofs, thresholds, specificities, sensitivities, concentrations of calibrators and / or controls etc.) used in an assay that employs specific sample type (e.g., such as an immunoassay that utilizes serum or a point-of-care device that employs whole blood) can be extrapolated to other assay formats using known techniques in the art, such as assay standardization. For example, one way in which assay standardization can be performed is by applying a factor to the calibrator employed in the assay to make the sample concentration read higher or lower to get a slope that aligns with the comparator method. Other methods of standardizing results obtained on one assay to another assay are wel known and have been described in the literature (See, for example, David Wild, Immunoassay Handbook, 4thedition, chapter 3.5, pages 315-322, the contents of which are herein incorporated by reference). As described herein, the assays and utilize monoclonal antibody 3H8 as a capture antibody (e.g., first specific binding partner) and monoclonal antibody 1A4, labeled with at least one detectable label, as a detection antibody (e.g., a second specific binding partner). In addition to the unique combination of capture and detection antibodies, the assays and methods also employ certain incubation and wash steps. As a result, the assaysand methodsof the present disclosure exhibit greater analytical sensitivity, namely, about a 3-fold lower limit of detection and about a 2-fold lower limit of quantification,when compared to the prior art methods, specificaly, the cMyC assay for use on the Erenna platform (Singulex). The cMyC assay for use on the Erenna platform utilizes monoclonal antibody 1A4 as the capture antibody and antibody 3H8 labeled with a detectable label, as the detection antibody. Additionaly, as discussed in Marjot, J., “The development and application of a high-sensitivity immunoassay for cardiac myosin-binding protein C,” Trans. Res., 170:17-25.eg (April 2016), the cMyC assay Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 optimized for use on the Erenna platform takes generates a result more than 3 hours after the start of the assay. Specificaly, in the assay optimized for use on the Erenna platform, microparticles coated with monoclonal antibody 1A4 are incubated with a sample for 2 hours. After a 2-hour incubation, monoclonal antibody 3H8, labeled with a detectable label, is added to the reaction mixture and incubated for 1 hour and then washed the result determined using single molecule counting. In addition to the improved analytical sensitivity, the assays and methods of the present disclosure also generate a result (e.g., the amount, level or concentration of cMyC) in less one hour, and in some instances, less than about 30 minutes from the start of the assay or method. In contrast, the cMyC assay for use on the Erenna platform generates a result in about 3hours from the start of the assay (See,Marjot, J., “The development and application of a high- sensitivity immunoassay for cardiac myosin-binding protein C,” Trans. Res., 170:17-25.eg (April 2016)).Other differences between the assays of the present invention and the assay optimized for use on the Erenna platform are provided in Table A, below.

[0002] Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 Table A Parameter Assay optimized for use on Assays and Methods of the Present th Ernn Pltfrm1 Dil r ardiac 3. Apparatus, Non-Transitory Machine-Readable and Computer Readable Storage Mediums, and Systems In some embodiments, disclosed herein are apparatus, machine-readable and computer readable storage mediums, and systems for use in implementing or performing the assays and methods described in Section 2. More specificaly, the apparatus, machine-readable and computer readable storage mediums and systems described herein can be used to identify cMyC in one or more biological samples obtained from a subject, determine the amount, concentration, or level of cMyC in one or more biological samples, and / orcommunicate from an apparatus (e.g., such as a point-of-care, non-point-of-care, or a point-of-care or non-point-of-care apparatus), whether the level of one or more biomarkers, is greater thanor less thanone or more reference or other levels. As used herein, the term “apparatus” is used interchangeably with “device” or “instrument”. In another embodiment, the present disclosure relates to an apparatus, device, or instrument. The apparatus, device, or instrument contains software to execute one or more tasks, including the performance of the methods described in Section2. In some embodiments, the apparatus, device, or instrumentcontains software to automaticaly determine the next Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 appropriate step in themethods described herein. For example, the apparatus, device, or instrumentmay contain software that determinesthe amount, concentration or level of cMyC. The software may display this determination, such as on a graphical user interface. In some embodiments, the apparatus, device, or instrumentsstores software that instructs processor or processor circuitryto execute or instantiate a given task. In some embodiments, the software stores machine-readable instructions that cause processor circuityto executeor instantiatea given task. The machine-readable instructions may be one or more executable programs or portion(s) of an executable program for execution by a computer. The programs may be embodied in software stored on a non-transitory computer readable storage medium such as a CD-ROM, a floppy disk, a hard drive, a DVD, a Blu-ray disk, or a memory associated with the processors. Alternatively, the entire programs and / or parts thereof could alternatively be executed by a device other than the processors and / or embodied in firmware or dedicated hardware. Additionaly,or alternatively, processes may be implemented by one or more hardware circuits (e.g., discrete and / or integrated analog and / or digital circuitry, an FPGA, an ASIC, a comparator, an operational-amplifier (op-amp), a logic circuit, etc.) structured to perform the coresponding operation without executing software or firmware. The machine-readable instructions may be stored in one or more of a compressed format, an encrypted format, a fragmented format, a compiled format, an executable format, a packaged format, etc. Machine readable instructions as described herein may be stored as data (e.g., portions of instructions, code, representations of code, etc.) that may be utilized to create, manufacture, and / or produce machine executable instructions. For example, the machine- readable instructions may be fragmented and stored on one or more storage devices and / or computing devices (e.g., servers). The machine-readable instructions may require one or more of instalation, modification, adaptation, updating, combining, supplementing, configuring, decryption, decompression, unpacking, distribution, reassignment, compilation, etc. in order to make them directly readable, interpretable, and / or executable by a computing device and / or other machine. For example, the machine-readable instructions may be stored in multiple parts, which are individualy compressed, encrypted, and stored on separate computing devices, wherein the parts when decrypted, decompressed, and combined form a set of executable instructions that implement a program such as that described herein. Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 In another example, the machine-readable instructions may be stored in a state in which they may be read by a computer, but require addition of a library (e.g., a dynamic link library (DLL), a software development kit (SDK), an application programming interface (API), etc. in order to execute the instructions on a particular computing device or other device. In another example, the machine-readable instructions may need to be configured (e.g., setings stored, data input, network addresses recorded, etc.)before the machine-readable instructions and / or the corresponding program(s) can be executed in whole or in part. Thus, the disclosed machine-readable instructions and / or coresponding program(s) are intended to encompass such machine-readable instructions and / or program(s) regardless of the particular format or state of the machine-readable instructions and / or program(s) when stored or otherwise at rest or in transit. The machine-readable instructions described herein can be represented by any past, present, or future instruction language, scripting language, programming language, etc. For example, the machine-readable instructions may be represented using any of the folowing languages: C, C++, Java, C#, Perl, Python, JavaScript, HyperText Markup Language (HTML), Structured Query Language (SQL), Swift, etc. The machine-readable instructions may be stored on a non-transitory computer and / or non-transitory machine-readable medium such as a hard disk drive, a flash memory, a read-only memory, a compact disk, a digital versatile disk, a cache, a random-access memory and / or any other storage device or storage disk in which information is stored for any duration (e.g., for extended time periods, permanently, for brief instances, for temporarily bufering, and / or for caching of the information). As used herein, the term “non-transitory computer readable medium”is defined to include any type of computer readable storage device and / or storage disk and to exclude propagating signals and to exclude transmission media. In some further embodiments, disclosed herein isasystemfor measuring or determining the amount, concentration, or level of cMyCin a biological sample.The system comprises: a. a data-obtaining module to obtaindataof an amount,concentration,or level of cMyC fromone or more assays performed on at least one or more biological samples obtained from a subject, where the one or more assays employ monoclonal antibody 3H8 as acapture Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 antibody and monoclonal antibody 1A4labeled with at least one detectable label as a conjugate or detection antibody; b. an evaluation module to analyze the data to obtain an evaluation result of the amount or level of cMyC in the biological samplewithin about 1 houror within about 30 minutesfrom the start of the assay(e.g., the start of the assay is when the biological sample is first contacted with either at least monoclonal antibody 3H8 or monoclonal antibody 1A4); and c.a data-outputingmodule to output the evaluation result. In further embodiments, disclosed hereinis a computer device that comprisesa storage device having machine-readable instructions stored thereon, and a processor.The processor executes the machine-readable instructionsto performthe steps of the above computer- implemented method,or the steps performed by the above evaluation system. In further embodiments, disclosed herein is an apparatus, device, or instrument that comprises a storage device having machine-readable instructions stored thereon, and a processor. The processor instantiatesor executesthe machine-readable instructions to: (A) identify cMyC in at least one or more biological samples obtained from a subject using at least one assay, where the at least one assay is performed using monoclonal antibody 3H8 as a capture antibody and monoclonal antibody 1A4labeled with at least one detectable label as a conjugate or detection antibody and the assay is completed within about 1 hour or about 30minutes from the start of the assay (e.g., the start of the assay is when the biological sample is first contacted with either at least monoclonal antibody 3H8 or monoclonal antibody 1A4); (B) determine an amount, concentration,orlevel of cMyCin one or more biological samples; and (C) communicatefrom the apparatus, device, or instrument(e.g., such as displaying on the apparatus, device, or instrument) the amount, concentration,or level of cMyC in the biological sample, or alternatively, whether the subject’s level of the cMyC is greater thanor less than a reference level forcMyC. In further embodiments, disclosed herein is a non-transitory machine-readable storage medium having machine-readable instructions stored thereon, wherein the machine-readable instructions are configured to be executed by a processor to perform the steps ofthe above Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 described computer-implemented method or the steps performed by the above one or more systems, such as the evaluation system. In yet further embodiments, disclosed herein is a non-transitory machine-readable storage medium. The non-transitory machine-readable storage medium can comprise instructions to cause one or processors or processor circuity to at least: (A) identify cMyC inat least one or more biological samples obtained from a subject using at least one assay,where the at least one assay is performed using monoclonal antibody 3H8 as a capture antibody and monoclonal antibody 1A4labeled with at least one detectable label as a conjugate or detection antibody and the assay is completed within about1 hour or about30minutes from the start of the assay (e.g., the start of the assay is when the biological sample is first contacted with either at least monoclonal antibody 3H8 or monoclonal antibody 1A4); (B) determine an amount, concentration, orlevel of cMyC in the one or more biological samples, determine a level of cMyC in one or more biological samples; and (C) communicatefrom an apparatus, device, or instrument(e.g., such as displaying on the apparatus, device, or instrument) the amount, concentration, or level of cMyC in one or more biological samples or whether the subject’slevel of cMyC is greaterthan or less than a reference level of cMyC. 4. Treatment and Monitoring of a Subject Sufering from Cardiac Injury The subject (e.g., a human subject) identified or assessed in the methods described above as having elevated levels of acMyC which of cardiac injury may be treated and / or monitored. In some embodiments, the method further includes treating the subject (e.g., human subject) determined as having elevated levels of cMyC with a cardiac injury treatment, usingany treatments known in the art. For example, such treatments can comprise administering to the subject: (a) one or more of a pharmaceutical agent (e.g., a thrombolytic or fibrinolytic drug (i.e., to break up a blood clot that is blocking blood flow to the heart), a blood thinneror anti- coagulant(e.g., aspirin, heparin, warfarin, clopidogrel, apixaban, dabigatran, dalteparin, edoxaban, enoxaparinor fondaparinux), nitroglycerin, morphine, beta blockers (e.g.,acebutolol, atenolol, betaxolol, bisoprolol / hydrochlorothiazide, bisoprolol, metoprolol, or nadolol), angiotensin-converting enzyme inhibitors(e.g., sulfhydryl-containing ACE inhibitors, dicarboxylic-containing ACE inhibitors, or phosphorus-containing ACE inhibitors, or a statin Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 (e.g, atorvastatin, fluvastatin, lovastatin, pitavastatin, pravastatin, rosuvastatin, or simvastatin); (b) oxygen; or (c) a combination of (a) and (b). Alternatively,or in addition, the method further includes performing one or more procedures on the subject. Such procedures can include one or moreof angioplasty, inserting of one or more stents, atherectomy, bypass surgery, or any combination thereof. In some embodiments, the method further includes monitoring the subject (e.g., a human subject) assessed as having elevated levels of cMyC. For example, in some embodiments the method further includes monitoring the subject assessed as having elevated levels of cMyC. In some embodiments, the subject having elevated levels of cMyC has not previously received any of the above-described treatments and / or procedures. In yet other embodiments, the subject having elevated levels of cMyC has previously receivedone or more of the above-described treatments and / or procedures. 5. Kit Provided herein is a kit, which may be used to determine the amount, concentration, or levelof cMyC in a biological sample. In some embodiments, the kit comprises monoclonal antibody 3H8 as a capture antibody and monoclonal antibody 1A4as the detection antibody. In yet further embodiments, kit can comprise instructions for assaying the test sample for cMyC by immunoassay, e.g., chemiluminescent microparticle immunoassay, a clinical chemistry assay, or any other assay known in the art(e.g., single molecule detection, lateral flow, or a point-of-care assay). Instructions included in kits can be afixed to packaging material or can be included as a package insert. While the instructions are typicaly writen or printed materials they are not limited to such. Any medium capable of storing such instructions and communicating them to an end user is contemplated by this disclosure. Such media include, but are not limited to, electronic storage media (e.g.,magnetic discs, tapes, cartridges, chips), optical media (e.g., CD ROM), and the like. As used herein, the term "instructions" can include the address of an internet site that provides the instructions.Alternatively or additionaly, the kit can comprise a calibrator or controlfor the at least one biomarker and / or atleast one container (e.g.,tube, microtiter plates or strips, which can be already coated with the relevant biomarker for conducting the assay, and / or a bufer, such as an assay bufer or a wash bufer, either one of which can be provided as a concentrated solution, a substrate solution for the detectable label (e.g.,an enzymatic label), or a stop solution. Preferably, the kit comprises al components, i.e., Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 reagents, standards, buffers, diluents, etc., which are necessary to perform the assay. The instructions also can include instructions for generating a standard curve. The kit may further comprise reference standards for quantifying cMyC. The reference standards may be employed to establish standard curves for interpolation and / or extrapolation of cMyC concentration. The reference standards may include a high cMyC concentration levelfor example, about 100000 pg / mL, about 125000 pg / mL, about 150000 pg / mL, about 175000 pg / mL, about 200000 pg / mL, about 225000 pg / mL, about 250000 pg / mL, about 275000 pg / mL, or about 300000 pg / mL; a medium cMyCconcentration level, for example, about 25000 pg / mL, about 40000 pg / mL, about 45000 pg / mL, about 50000 pg / mL, about 55000 pg / mL, about 60000 pg / mL, about 75000 pg / mL or about 100000 pg / mL; and / or a low cMyC concentration level, for example, about 1 pg / mL, about 5 pg / mL, about 10 pg / mL, about 12.5 pg / mL, about 15 pg / mL, about 20 pg / mL, about 25 pg / mL, about 30 pg / mL, about 35 pg / mL, about 40 pg / mL, about 45 pg / mL, about 50 pg / mL, about 55 pg / mL, about 60 pg / mL, about 65 pg / mL, about 70 pg / mL, about 75 pg / mL, about 80 pg / mL, about 85 pg / mL, about 90 pg / mL, about 95 pg / mL, or about 100 pg / mL.In some embodiments, the reference standard for cMyC concentration is 0. Any antibodies, which are provided in the kit, such as monoclonal or recombinant antibodies specific for cMyC(e.g., monoclonal antibody 1A4), can incorporate a detectable label, such as a fluorophore, radioactive moiety, enzyme, biotin / avidin label, chromophore, chemiluminescent label, or the like, or the kit can include reagents for labeling the antibodies or reagents for detecting the antibodies (e.g., detection antibodies) and / or for labeling cMyC. The antibodies, calibrators, and / or controls can be provided in separate containers or pre-dispensed into an appropriate assay format, for example, into microtiter plates. Optionaly, the kit includes quality control components (for example, sensitivity panels, calibrators, and positive controls). Preparation of quality control reagents is wel-known in the art and is described on insert sheets for a variety of immunodiagnostic products. Sensitivity panel members are optionalyused to establish assay performance characteristics, and further optionaly are useful indicators of the integrity of the immunoassay kit reagents, and the standardization of assays. The kit can also optionaly include other reagents required to conduct a diagnostic assay or facilitate quality control evaluations, such as bufers, salts, enzymes, enzyme co-factors, Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 substrates, detection reagents, and the like. Other components, such as buffers and solutions for the isolation and / or treatment of a test sample (e.g., pre-treatment reagents), also can be included in the kit. The kit can additionaly include one or more other controls. One or more of the components of the kit can be lyophilized, in which case the kit can further comprise reagents suitable for the reconstitution of the lyophilized components. The various components of the kit are optionalyprovided in suitable containers as necessary, e.g., a microtiter plate. The kit can further include containers for holding or storing a sample (e.g., a container or cartridge for a urine, whole blood, plasma, or serum sample). Where appropriate, the kitoptionaly also can contain reaction vessels, mixing vessels, and other components that facilitate the preparation of reagents or the test sample. The kit can also include one or more instrumentsfor assisting with obtaining a test sample, such as a syringe, pipete, forceps, measured spoon, or the like. If the detectable label is at least one acridinium compound, the kit can comprise at least one acridinium-9-carboxamide, at least one acridinium-9-carboxylate aryl ester, or any combination thereof. If the detectable label is at least one acridinium compound, the kit also can comprise a source of hydrogen peroxide, such as a bufer, solution, and / or at least one basic solution. If desired, the kit can contain a solid phase, such as a magnetic particle, bead, test tube, microtiter plate, cuvete, membrane,scafolding molecule, film, filter paper, disc, or chip. If desired, the kit can further comprise one or more components, alone or in further combination with instructions, for assaying the test sample for another analyte, which can be a biomarker, such as a biomarker ofcardiac injury. 6. Adaptation of Kit and Method The kit (or components thereof), as wel as the method for assessing or determining the concentration of cMyC in a test sample by an immunoassay as described herein, can be adapted for use in a variety of automated and semi-automated systems (including those wherein the solid phase comprises a microparticle), as described, e.g., U.S. Patent No.5,063,081, U.S. Patent Application Publication Nos.2003 / 0170881, 2004 / 0018577, 2005 / 0054078, and 2006 / 0160164 and as commercialy marketed e.g., by Abbot Laboratories (Abbot Park, IL) as Abbot Point of Care (i-STAT or i-STAT Alinity, Abbot Laboratories) as wel as those described in U.S. Patent Nos.5,089,424 and 5,006,309, and as commercialy marketed, e.g.,by Abbot Laboratories (Abbot Park, IL) as ARCHITECT® or the series of Abbot Alinity devices. Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 Some of the diferences between an automated or semi-automated system as compared to a non-automated system (e.g., ELISA) include the substrate to which the first specific binding partner (e.g., analyte antibody or capture antibody) is atached (which can afect sandwich formation and analyte reactivity), and the length and timing of the capture, detection, and / or any optional wash steps. Whereas a non-automated format such as an ELISA may require a relatively longer incubation time with sample and capturereagent (e.g., about 2 hours), an automated or semi-automated format (e.g.,ARCHITECT®, Alinity, and any successor platform, Abbot Laboratories) may have a relatively shorter incubation time (e.g.,approximately 18 minutes for ARCHITECT®). Similarly, whereas a non-automated format such as an ELISA may incubate a detection antibody such as the conjugate reagent for a relatively longer incubation time (e.g.,about 2 hours), an automated or semi-automated format (e.g., ARCHITECT®, Alinity, and any successorplatform) may have a relatively shorter incubation time (e.g., approximately 4 minutes for the ARCHITECT® and any successor platform). Other platforms available from Abbot Laboratories include, but are not limited to, Alinity, AxSYM®, IMx® (see, e.g., U.S. Patent No.5,294,404, which is hereby incorporated by reference in its entirety), PRISM®, EIA (bead), and Quantum™I, as wel as other platforms. Additionaly, the assays, kits, and kit components can be employed in other formats, for example, on electrochemical or other hand-held or point-of-care assay systems. As mentioned previously, the present disclosure is, for example, applicable to the commercial Abbot Point of Care (i-STAT®, Abbot Laboratories) electrochemical immunoassay system that performs sandwich immunoassays. Immunosensors and their methods of manufacture and operation in single-use test devices are described, for example in, U.S. Patent No.5,063,081, U.S. Patent App. Publication Nos.2003 / 0170881, 2004 / 0018577, 2005 / 0054078, and 2006 / 0160164, which are incorporated in their entireties by reference for their teachings regarding same. Regarding the adaptation of an assay to the i-STAT system, the folowing configuration is prefered. A microfabricated silicon chip is manufactured with a pair of gold amperometric working electrodes and a silver-silver chloride reference electrode. On one of the working electrodes, polystyrene beads (0.2 mm diameter) with immobilized capture antibody are adhered to a polymer coating of paterned polyvinyl alcohol over the electrode. This chip is assembled into an i-STAT® cartridge with a fluidics formatsuitable for immunoassay. On a portion of the silicon chip, there is a specific binding partnerfor cMyC, cMyC antibodies (one or Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 more monoclonal / polyclonal antibody or a fragment thereof, a variant thereof, or a fragment of a variant thereof that can bindcMyC) or a fragment of a variant thereof that can bindcMyC), either of which can be detectably labeled. Within the fluid pouch of the cartridge is an aqueous reagent that includes p-aminophenol phosphate. In operation, a sample from a subject suspected of sufering from a cardiac injuryis added to the holding chamber of the test cartridge, and the cartridge is inserted into the i-STAT reader. A pump element within the cartridge pushes the sample into a conduit containing the chip. The sample is brought into contact with the sensors alowing the enzyme conjugate to dissolve into the sample. The sample is oscilated across the sensors to promote formation of the sandwich of approximately 2-12 minutes. In thepenultimate step of the assay, the sample is pushed into a waste chamber and wash fluid, containing a substrate for the alkaline phosphatase enzyme, is used to wash excess enzyme conjugate and sample of the sensor chip. In the final step of the assay, the alkaline phosphatase label reacts with p-aminophenol phosphate to cleave the phosphate group and permit the liberated p-aminophenol to be electrochemicaly oxidized at the working electrode. Based on the measured current, the reader is able to calculatethe amount of cMyC in the sample by means of an embedded algorithm and factory-determined calibration curve. The methods and kits as described herein necessarily encompass other reagents and methods for carying out the immunoassay. For instance, encompassed are various bufers such as are known in the art and / or which can be readily prepared or optimized to be employed, e.g., for washing, as a conjugate diluent, and / or as a calibrator diluent. An exemplary conjugate diluent is ARCHITECT conjugate diluent employed in certain kits (Abbot Laboratories, Abbot Park, IL) and containing 2-(N-morpholino)ethanesulfonic acid (MES), a salt, a protein blocker, an antimicrobial agent, and a detergent. An exemplary calibrator diluent is ARCHITECT human calibrator diluent employed in certain kits (Abbot Laboratories, Abbot Park, IL), which comprises a buffer containing MES, other salt, a protein blocker, and an antimicrobial agent. Additionaly, as described in U.S. Patent Application No.61 / 142,048 filed December 31, 2008, improved signal generation may be obtained, e.g.,in an i-STAT cartridge format, using a nucleic acid sequence linked to the signal antibody as a signal amplifier. The present disclosure has multiple embodiments, ilustrated by the folowing non- limiting examples. Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 EXAMPLES The folowing examples are for the purposesof ilustration only and are not intended tolimitthe scope ofthe claims. EXAMPLE 1 Cardiac Myosin Binding Protein C (cMyC) Immunoassay 150 uL human sample (serum or plasma), 50 uL of reagent containing acridinylated anti-cMyC antibody, and 50 uL of reagent containing anti-cMyC coated microparticles were added to a reaction vessel, vortexed, and incubated for 18 minutes (See, FIG.1). Folowing this incubation, the microparticles were sequestered at the side of the reaction vessel using a magnet while the reaction supernatant was removed. The microparticles are subsequently washed with wateror detergent solution(colectively “Wash Solution”).During this initial step, the anti- cMyC antibody-acridinium conjugate: cMyCantigen: anti-cMyCcoated microparticle complexes are formed and captured by the microparticles. In the second step, immediately folowing washing, 50 uL of a wash bufer containing 2-(N-morpholino)ethanesulfonic acid (MES), a salt, a protein blocker, an antimicrobial agent, and a detergent,was added to the reaction vessel, vortexed and alowed to incubate for 4 minutes. During this step the microparticles and bound complex are washed from other non-specificaly binding components. Folowing incubation, the microparticles were sequestered at the side of the reaction vessel using a magnet and the reaction supernatant removed. The microparticles were subsequently washed witha Wash Solution. Washed particles were suspended in a basic-hydrogen peroxide containing solution to activate the acridinium with simultaneous measurement of light output (in relative light units or RLU), which is proportional to the amount of conjugate bound onto the microparticles. The detection limits of the above-described assay are shown below in Table 1 and in FIG.3. Table 1 cMyc Concentration (ng / L) % CV Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 3.0 3.20 4.0 3.34 Additionaly, the linearity of the above-described assay is shown below in Table 2 and in FIG.4. Table 2 Observed cMyC Expected cMyC Concentration % Diference Concentration (ng / L) (ng / L) The above results demonstrate a linearity with ± 15% between 5 to 6800 ng / L of cMyC observed. Finaly, FIG.5 shows the detection of cMyC in anassumed normal, healthy population from subjects donating blood in the U.S.using the above assay. FIG.6 shows the distribution of cMyC concentrations in random populations of samples obtained from subjects having elevated TnI. Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 It is understood that the foregoing detailed description and accompanying examples are merely ilustrative and are not to be taken as limitations upon the scope of the disclosure, which is defined solely by the appended claims and their equivalents. Various changes and modifications to the disclosed embodiments wil be apparent to those skiled in the art. Such changes and modifications, including without limitation those relating to the chemical structures, substituents, derivatives, intermediates, syntheses, compositions, formulations, or methods of use of the disclosure, may be made without departing from the spirit and scope thereof. For reasons of completeness, various aspects of the disclosure are set out in the folowing numbered clauses: Clause 1. A method comprising: a) receiving a level of cardiac myosin binding protein C (“cMyC”) from an assay performed on at least one biological sample obtained from a subject; and b) determining whether the level of cMyC obtained in step a) is higher or lower than a reference level, wherein: i. if the level of cMyC determined in step b) is: (A) higher than the reference level, further determining that the subject is sufering from a cardiac injury; or (B) lower than the reference level, further determining that the subject is not sufering from acardiac injury; and i. performing the assay comprises: A)contacting the biological sample either simultaneously or sequentialy, in any order, with: at least one first specific binding partner atached to a solid support, wherein the at least one first specific binding partner comprises monoclonal antibody 3H8 which specificaly binds to at least one epitope on cMyC, and at least one second specific binding partner comprising a detectable label, wherein the at least one second specific binding partner comprises monoclonal antibody 1A4 which specificaly binds to at least one epitope on cMyC which is a diferent epitope than the epitope bound by monoclonal antibody 3H8, to form a first mixture, Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 B) incubating the first mixture for about 12 to about 30 minutes to form one or more first complexes comprising the first specific binding partner-cMyC-second specific binding partner; C) isolating the first specific binding partner-cMyC-second specific binding partner complexes from the first mixture; D) washing the first specific binding partner-cMyC-second specific binding partner complex of step C) with a solution comprising water or at least one detergent to form a second mixture comprising the first specific binding partner-cMyC-second specific binding partner; E) incubating the second mixture comprising the first specific binding partner-cMyC- second specific binding partner complex for about 1 to about 5 minutes; F) isolating the first specific binding partner-cMyC-second specific binding partner complexes from the second mixture; G) washing the isolated first specific binding partner-cMyC-second specific binding partner complex of step F) with a solution comprising water or at least one detergent; and H) assessing a signal from the one or more first complexes, wherein the amount of detectable signal from the detectable label indicates the presence or amount of cMyC in the sample. Clause 2. The method of clause1, wherein, the biological sample is a whole blood, serum, or plasma sample. Clause 3. The method of clause1 or clause2, wherein the solid support is a microparticle. Clause 4. The method of any of clauses1-3, wherein the cardiac injury myocardial infarction or reinfarction. Clause 5. The method of any of clauses1-4, wherein antibody 3H8 is produced by hybridoma cel lineDSM ACC3223. Clause 6.The method of any of clauses1-5, wherein antibody 1A4 is produced by hybridoma cel line DSM ACC3224. Clause 7. The method of any of clauses1-6, wherein the method comprises determining that the subject is sufering from a cardiac injury. Clause 8. The method of clause7, wherein the cardiac injury is myocardial infarction or reinfarction. Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 Clause 9. The method of clause 8, wherein the myocardial infarction is acute myocardial infarction. Clause 10. The method of any of clauses7-9, wherein the method further comprises treating the patient for the cardiac injury, wherein the treatment comprises: (a) administering to the subject one or more pharmaceutical agents, oxygen, or a combination thereof; (b) performing one or more procedures on the subject, wherein said procedures are angioplasty, inserting a stent, atherectomy, bypass surgery, or combinations thereof; or (c) any combination of (a) and (b). Clause 11. The method of any of clauses7-10, where the method further comprises monitoring a subject sufering a cardiac injury. Clause 12. The method of any of clauses1-11, wherein the assay is an immunoassay or a clinical chemistry assay. Clause 13. The method of any of clauses1-12, wherein the assay is performed using single molecule detection, lateral flow, or a point-of care method. Clause 14. The method of any of clause1-13, wherein the first mixture is incubated for about 18 minutes. Clause 15. The method of any of clause1-14, wherein the second mixture is incubated for about 4 minutes. Clause 16. A method comprising: a) receiving a level of cardiac myosin binding protein C (“cMyC”) from an assay performed on at least one biological sample obtained from a subject; and b) determining whether the level of cMyC obtained in step a) is higher or lower than a reference level, wherein: i. if the level of cMyC determined in step b) is: (A) higher than the reference level, further determining that the subject is sufering from a cardiac injury; or (B) lower than the reference level, further determining that the subject is not sufering from a cardiac injury; and i. performing the assay comprises: A)contacting the biological sample either simultaneously or sequentialy, in any order, with: Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 at least one first specific binding partner atached to a solid support, wherein the at least one first specific binding partner comprises monoclonal antibody 3H8 which specificaly binds to at least one epitope on cMyC, and at least one second specific binding partner comprising a detectable label, wherein the at least one second specific binding partner comprises monoclonal antibody 1A4 which specificaly binds to at least one epitope on cMyC which is a diferent epitope than the epitope bound by monoclonal antibody 3H8, to form a first mixture, B) incubating the first mixture for about 18minutes to form one or more first complexes comprising the first specific binding partner-cMyC-second specific binding partner; C) isolating the first specific binding partner-cMyC-second specific binding partner complexes from the first mixture; D) washing the first specific binding partner-cMyC-second specific binding partner complex of step C) with a solution comprising water or at least one detergent to form a second mixture comprising the first specific binding partner-cMyC-second specific binding partner; E) incubating the second mixture comprising the first specific binding partner-cMyC- second specific binding partner complex for about 4minutes; F) isolating the first specific binding partner-cMyC-second specific binding partner complexes from the second mixture; G) washing the isolated first specific binding partner-cMyC-second specific binding partner complex of step F) with a solution comprising water or at least one detergent; and H) assessing a signal from the one or more first complexes, wherein the amount of detectable signal from the detectable label indicates the presence or amount of cMyC in the sample. Clause 17. The method of clause 16, wherein the biological sample is a whole blood, serum, or plasma sample. Clause 18. The method of clause 16, wherein the biological sample is a whole blood sample. Clause 19. The method of clause 16, wherein the biological sample is a serum sample. Clause 20. The method of clause 16, wherein the biological sample is a plasma sample. Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 Clause 21. The method of any of clauses 16-20, wherein the solid support is a microparticle. Clause 22. The method of any of clauses16-21, wherein the cardiac injury myocardial infarction or reinfarction. Clause 23. The method of any of clauses16-22wherein antibody 3H8 is produced by hybridoma cel lineDSM ACC3223. Clause 24.The method of any of clauses16-23, wherein antibody 1A4 is produced by hybridoma cel line DSM ACC3224. Clause 25. The method of any of clauses 16-24, wherein the method comprises determining that the subject is sufering from a cardiac injury. Clause 26. The method of clause 24, wherein the cardiac injury is myocardial infarction or reinfarction. Clause 27. The method of clause 26, wherein the myocardial infarction is acute myocardial infarction. Clause 28. The method of any of clauses25-27, wherein the method further comprises treating the patient for the cardiac injury, wherein the treatment comprises: (a) administering to the subject one or more pharmaceutical agents, oxygen, or a combination thereof; (b) performing one or more procedures on the subject, wherein said procedures are angioplasty, insertinga stent, atherectomy, bypass surgery, or combinations thereof; or (c) any combination of (a) and (b). Clause 29. The method of any of clauses25-27, where the method further comprises monitoring a subject sufering a cardiac injury. Clause 30. The method of any of clauses16-29, wherein the assay is an immunoassay or a clinical chemistry assay. Clause 31. The method of any of clauses 16-29, wherein the assay is an immunoassay. Clause 32. The method of any of clauses 16-29, wherein the assay is a clinical chemistry assay. Clause 33. The method of any of clauses 16-32, wherein the assay is performed using single molecule detection, lateral flow, or a point-of care method. Clause 34. The method of any of clause 16-32, wherein the assay is performed using single molecule detection. Client Ref No.15766WOO1 Aty Docket No. ABBTL-43031.601 Clause 35. The method of any of clauses 16-32, wherein the assay is performed using a lateral flow method. Clause 36. The method of any of clauses 16-32, wherein the assay is performed using a point-of-care method. Clause 37. The method of any of clauses 1-15, wherein the method is quantitative. Clause 38. The method of any of clauses 1-15, wherein the method is qualitative.

Claims

Client Ref No.15766WOO1 Aty Docket No. ABBTL- 43031.601 CLAIMS We claim:

1. A method comprising: a) receiving a level of cardiac myosin binding protein C (“cMyC”) from an assay performed on at least one biological sample obtained from a subject; and b) determining whether the level of cMyC obtained in step a) is higher or lower than a reference level, wherein: i. if the level of cMyC determined in step b) is: (A) higher than the reference level, further determining that the subject is sufering from a cardiac injury; or (B) lower than the reference level, further determining that the subject is not sufering from a cardiac injury; and i. performing the assay comprises: A) contacting the biological sample either simultaneously or sequentialy, in any order, with: at least one first specific binding partner atached to a solid support, wherein the at least one first specific binding partner comprises monoclonal antibody 3H8 which specificaly binds to at least one epitope on cMyC, and at least one second specific binding partner comprising a detectable label, wherein the at least one second specific binding partner comprises monoclonal antibody 1A4 which specificaly binds to at least one epitope on cMyC which is a diferent epitope than the epitope bound by monoclonal antibody 3H8, to form a first mixture, B) incubating the first mixture for about 12 to about 30 minutes to form one or more first complexes comprising the first specific binding partner-cMyC-second specific binding partner; C) isolating the first specific binding partner-cMyC-second specific binding partner complexes from the first mixture;Client Ref No.15766WOO1 Aty Docket No. ABBTL- 43031.601 D) washing the first specific binding partner-cMyC-second specific binding partner complex of step C) with a solution comprising water or at least one detergent to form a second mixture comprising the first specific binding partner-cMyC-second specific binding partner; E) incubating the second mixture comprising the first specific binding partner-cMyC-second specific binding partner complex for about 1 to about 5 minutes; F) isolating the first specific binding partner-cMyC-second specific binding partner complexes from the second mixture; G) washing the isolated first specific binding partner-cMyC-second specific binding partner complex of step F) with a solution comprising water or at least one detergent; and H) assessing a signal from the one or more first complexes, wherein the amount of detectable signal from the detectable label indicates the presence or amount of cMyC in the sample.

2. The method of claim 1, wherein, the biological sample is a whole blood, serum, or plasma sample.

3. The method of claim 1 or claim 2, wherein the solid support is a microparticle.

4. The method of any of claims 1-3, wherein the cardiac injury myocardial infarction or reinfarction.

5. The method of any of claims 1-4, wherein antibody 3H8 is produced by hybridoma cel line DSM ACC3223.

6. The method of any of claims 1-5, wherein antibody 1A4 is produced by hybridoma cel line DSM ACC3224.

7. The method of any of claims 1-6, wherein the method comprises determining that the subject is sufering from a cardiac injury.

8. The method of claim 7, wherein the cardiac injury is myocardial infarction or reinfarction.

9. The method of claim 8, wherein the myocardial infarction is acute myocardial infarction.

10. The method of any of claims 7-9, wherein the method further comprises treating the patient for the cardiac injury, wherein the treatment comprises: (a) administering to the subjectClient Ref No.15766WOO1 Aty Docket No. ABBTL- 43031.601 one or more pharmaceutical agents, oxygen, or a combination thereof; (b) performing one or more procedures on the subject, wherein said procedures are angioplasty, inserting a stent, atherectomy, bypass surgery, or combinations thereof; or (c) any combination of (a) and (b).

11. The method of any of claims 7-10, where the method further comprises monitoring a subject sufering a cardiac injury.

12. The method of any of claims 1-11, wherein the assay is an immunoassay or a clinical chemistry assay.

13. The method of any of claims 1-12, wherein the assay is performed using single molecule detection, lateral flow, or a point-of care method.

14. The method of any of claims 1-13, wherein the first mixture is incubated for about 18 minutes.

15. The method of any of claims 1-14, wherein the second mixture is incubated for about 4 minutes.

16. The method of any of claims 1-15, wherein the method is quantitative.

17. The method of any of claims 1-15, wherein the method is qualitative.

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