CN-121991220-A - Antibody for detecting cartilage oligomeric matrix protein, kit and application

Abstract

The application discloses an antibody for detecting cartilage oligomeric matrix protein, a kit and application thereof. The antibody is prepared by immunizing animals after coupling cartilage oligomeric matrix protein specific epitope peptide with carrier protein, and the amino acid sequence of the cartilage oligomeric matrix protein specific epitope peptide is at least one of a sequence shown as SEQ ID NO.1 and a sequence shown as SEQ ID NO. 2. The antibody can specifically detect cartilage oligomeric matrix protein, has the advantages of high sensitivity, good specificity, good repeatability and the like, has good diagnosis sensitivity, specificity and repeatability when being used for auxiliary diagnosis or curative effect evaluation of osteoarthritis diseases, meets the clinical application requirements, provides a more accurate and reliable objective index for auxiliary diagnosis, disease condition monitoring and prognosis evaluation of related diseases, and has important significance and clinical application value.

Inventors

• ZHU JIANAN
• LI YUPING
• SHENG JIAHUA
• WANG ZHENYU
• GU HUIXIAN
• LIU YAO

Assignees

• 深圳市安群生物工程有限公司
• 深圳市药品检验研究院（深圳市医疗器械检测中心）

Dates

Publication Date

20260508

Application Date

20251231

Claims (10)

1. 1. An antibody for detecting cartilage oligomeric matrix protein is characterized in that the antibody is prepared by immunizing animals after coupling a cartilage oligomeric matrix protein specific antigen epitope peptide with carrier protein, and the amino acid sequence of the cartilage oligomeric matrix protein specific antigen epitope peptide is at least one of a sequence shown as SEQ ID NO.1 and a sequence shown as SEQ ID NO. 2; SEQ ID NO.1: Tyr-Asp-Val-Arg-Glu-Leu-Leu-Arg-Gln-Gln-Val-Arg-Glu-Ile-Thr SEQ ID NO.2: Tyr-Asp-Gln-Ala-Asp-Val-Asp-His-Asp-Phe-Val-Gly-Asp-Ala。
2. 2. the antibody according to claim 1, wherein the antibody is a polyclonal antibody prepared by immunizing an animal after the cartilage oligomeric matrix protein specific antigen epitope peptide is coupled with a carrier protein.
3. 3. A kit for detecting cartilage oligomeric matrix protein, which is characterized by comprising the antibody of claim 1 or 2.
4. 4. The kit of claim 3, comprising a solid support and said antibody, wherein said antibody is directly or indirectly attached to said solid support; Optionally, the solid support is at least one of a microporous reaction plate, magnetic microspheres, and nitrocellulose membrane.
5. 5. The kit according to claim 4, further comprising a label for labeling the antibody; Optionally, the label is at least one of horseradish peroxidase HRP, alkaline phosphatase AP, a luminescent substance, a fluorescent substance, a dye, and colloidal gold.
6. 6. The kit according to claim 4, further comprising a cartilage oligomeric matrix protein standard.
7. 7. The kit according to any one of claims 3 to 6, wherein the kit is at least one of an ELISA kit, a chemiluminescent assay kit, a fluorescent immunochromatographic assay kit and a colloidal gold immunoassay kit.
8. 8. The kit according to claim 7, wherein the chemiluminescent assay kit is prepared by combining a chemiluminescent immunoassay with a double antibody sandwich method.
9. 9. The kit of claim 7, further comprising a magnetic particle suspension, a wash solution, a chemiluminescent substrate; Optionally, the chemiluminescent substrate comprises at least one of luminol, isoluminol and derivatives thereof, (adamantane) -1, 2-dioxyethane and derivatives thereof.
10. 10. Use of the specific antibody of claim 1 or 2, or the kit of any one of claims 3-9, in the manufacture of a product for diagnosing osteoarthritis.

Description

Antibody for detecting cartilage oligomeric matrix protein, kit and application Technical Field The application relates to the technical field of protein detection, in particular to an antibody for detecting cartilage oligomeric matrix protein, a kit and application thereof. Background Cartilage Oligomeric Matrix Protein (COMP) is a glycoprotein in the form of a "bouquet-like" pentameric structure in articular cartilage extracellular matrix (ECM) assembled from 5 524 kDa monomers with a C-terminal oligomerization domain via a coiled coil. The method is similar to a 'molecular rivet', and the II type collagen fiber and the aggrecan (aggrecan) are firmly riveted together to construct a three-dimensional net structure which is compression resistant and extremely elastic, so that the normal joint still keeps smooth and low friction in millions of load cycles. When chondrocytes are subjected to an inflammatory, mechanical overload or metabolic imbalance stress to initiate a "suicide" program, proteases such as MMP-13, ADAMTS-4/5, etc., will cleave precisely like "molecular scissors" in the T3 repeat region of COMP, yielding characteristic fragments such as 110 kDa, 85 kDa, etc. After cleavage by proteases, the COMP fragments together with the collagen fibrillar fragments escape into the joint space, forming a soluble osteoarthritis product. Since COMP has a half-life in the systemic circulation of only 6-12 h, its serum or synovial concentration can be significantly increased within hours and reflect the breakdown kinetics of the cartilage matrix instantaneously with high temporal resolution, it is considered as one of the currently most sensitive dynamic biomarkers of osteoarthritis. However, as cartilage matrix degradation events continue to accumulate in the individual microenvironment, their pathological signals, after epidemiological scale-up, have evolved into a systemic crisis with significant public health and economic overflow effects worldwide. World health organization Global HEALTH ESTIMATES 2023 indicates that Osteoarthritis (OA) is 5.28 hundred million and Rheumatoid Arthritis (RA) is about 1.80 hundred million, and that the age-standardized prevalence of cartilage degeneration-related diseases in 50-79 year old people is greater than 15% if post-traumatic and inflammatory spinal arthropathy are incorporated. The pain and dysfunction directly contributes to medical expenditure, the US medical expenditure database (MEPS 2022) shows that the annual cost of OA is directly 1,360 hundred million dollars (including joint cavity injection, joint replacement and hospitalization), RA and its complications are increased by 380 hundred million dollars, and the European health and economy alliance (EU-HEA 2022) reports an overall economic burden of OA of 2,400 hundred million Euro accounting for 2.7% of the overall European health expenditure. On the labor level, OA patients on average leave the market 2.8 years earlier, RA years are absenteeism 35 d (IQR 28-42 d), and U.S. productivity losses (early retirement, absenteeism, post replacement, and human capital trade-off) have exceeded 1,800 billion dollars, accounting for 0.7% of their GDP. Chronic disability further induces co-diseases such as depression, cardiovascular diseases, type 2 diabetes and the like, so that the health-related quality of life (HRQoL) is reduced by 35-60%, the injury and residue caused by cartilage degeneration worldwide in 2022 is regulated to be as high as 4,900 thousands of years, and the indirect social cost is exponentially increased. From this, it can be seen that the instantaneous release of COMP fragments in the joint cavity not only marks the microscopic disintegration of the cartilage matrix, but also accumulates in the global hygienically economical level into several trillions of dollars of direct medical expenditure and indirect productivity loss through the disease burden of cascade amplification, which affects beyond the molecular event itself, becoming a systematic risk that is difficult to carry by public health systems and macro-economic structures. COMP can be summarized as a four-level network of structural scaffolds-mechanical homeostasis-signal junction-pathological amplification. (1) The structural layer, COMP, through the T3 repetitive region and Ca 2+ -dependent manner cross-links with the notch region of the type II collagen fiber, so that the tensile strength of the collagen network is improved by 20-30%, the compressive modulus is improved by about 1.5 times, and the EGF-like structural domain is combined with the hydroxyapatite crystal surface in the cartilage of the growth plate to block the active site of crystal growth and inhibit abnormal calcification (Ki is about 5.4 mu M). (2) On the cellular level, COMP and integrin alpha 5 beta 1 are combined to trigger FAK-Y397-PI 3K-p 85-AKT-S473-mTorrC 1 cascade, promote cartilage cell proliferation and inhibit caspase-3 mediated apoptosis, and in inflammatory