Proteolytic markers as diagnostic biomarkers for cancer, organ injury and muscle rehabilitation/exercise overtraining
View Patent ↗The present invention identifies biomarkers that are diagnostic of nerve cell injury, organ injury, and/or neuronal disorders. Detection of different biomarkers of the invention are also diagnostic of the degree of severity of nerve injury, the cell(s) involved in the injury, and the subcellular localization of the injury.
1. A kit for detecting muscle group stress or injury in a subject,
the kit comprising:
(a) a substrate for holding a biological sample isolated from a human subject suspected of having a muscle group injury or stress;
(b) at least one isolated fragment-specific antibody or a plurality thereof that specifically and independently binds to a calpain or caspase-3 induced breakdown product of a troponin protein, wherein the troponin protein breakdown products are breakdown fragments of troponin, troponin C, troponin C-slow skeletal/cardiac (TN-C), skeletal troponin C-fast (STNC), troponin T, troponin T1, T2, T3, troponin t, troponin I1, I2, I3 or mixtures thereof; and
(c) printed instructions for reacting the isolated fragment-specific antibody with the biological sample or a portion of the biological sample to detect the presence or amount of a troponin protein breakdown fragment in the biological sample, wherein said troponin breakdown fragments are detected with said antibodies prepared against the cleavage sites of troponin selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 42, SEQ ID NO: 43, and SEQ ID NO: 44 or mixtures thereof which are present in biological samples of a subject having cardiac muscle injury, and wherein said troponin breakdown fragments are absent in a normal subject not having cardiac muscle injury; or
wherein said troponin breakdown fragments in a normal subject are present in an amount less than 20% of the troponin breakdown fragments found in a subject having smooth or skeletal muscle injury or muscle stress.
2. The kit of claim 1 wherein the biological fluid sample is whole blood, serum, plasma, cerebral spinal fluid (CSF), sweat or saliva.
3. The kit of claim 1 wherein the skeletal or smooth muscle injury or stress is identified with spontaneous or drug induced muscle waste, rhabdomyolysis, muscular dystrophy, cardiac stress, heart failure, cardiac ischemia, myocardial infarction, muscle or exercise overtraining.
4. The kit of claim 1 wherein the kit is used with an immunoassay to react the isolated fragment-specific antibody with the biological sample or a portion of the biological sample to detect the presence or amount of a troponin protein breakdown fragment in the biological sample.
5. The kit of claim 4 wherein the immunoassay is an Enzyme-linked immunosorbent assay (ELISA).
6. The kit of claim 5 wherein the ELISA is a sandwich assay.
7. The kit of claim 1 wherein said specific and independently binding isolated fragment-specific antibodies are prepared against N-terminus or C-terminus epitopes of troponin, troponin C, troponin C-slow TN-C, STNC, troponin T, troponin T1, T2, T3, troponin I, troponin I1, I2, or I3 selected from the group consisting of SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO 55, SEQ ID NO: 56, SEQ ID NO 57, SEQ ID NO: 58, SEQ ID NO 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO: 134, SEQ ID NO: 135, and SEQ ID NO: 136.