WO-2026091296-A1 - CARDIAC MYOSIN-BINDING PROTEIN C DETECTION KIT AND USE THEREOF

WO2026091296A1WO 2026091296 A1WO2026091296 A1WO 2026091296A1WO-2026091296-A1

Abstract

Provided herein is a myosin binding protein C detection kit, comprising: 1) a magnetic bead coupled with a first antibody, and 2) a second antibody coupled with an acridinium ester, the first antibody and the second antibody both being human myosin binding protein C antibodies, but binding to different antigenic epitopes. By means of a double antibody sandwich method and the chemiluminescence characteristics of the acridinium ester, the myosin binding protein C detection kit detects the content of myosin binding protein C in serum with high sensitivity.

Inventors

GE, JUNBO
CHENG, Leilei
WANG, YAN
CAI, Qingqing
CHEN, HUIYONG
ZHANG, SHILONG
LIU, Rongle
XU, Yuchen
LIU, YUAN

Assignees

复旦大学附属中山医院

Dates

Publication Date: 20260507
Application Date: 20241231
Priority Date: 20241028

Claims (4)

The myosin-binding protein C assay kit includes: 1) Magnetic beads conjugated with a first antibody, wherein the heavy chain variable region of the first antibody includes HCDR1 (SEQ ID NO:3), HCDR2 (SEQ ID NO:4), and HCDR3 (SEQ ID NO:5), and the light chain variable region of the first antibody includes LCDR1 (SEQ ID NO:7), LCDR2 (SEQ ID NO:8), and LCDR3 (SEQ ID NO:9); and 2) A second antibody conjugated with acridinium ester, wherein the heavy chain variable region of the second antibody includes HCDR1 (SEQ ID NO: 11), HCDR2 (SEQ ID NO: 12), and HCDR3 (SEQ ID NO: 13), and the light chain variable region of the first antibody includes LCDR1 (SEQ ID NO: 15), LCDR2 (SEQ ID NO: 16), and LCDR3 (SEQ ID NO: 17). or 1) Magnetic beads conjugated with a first antibody, wherein the heavy chain variable region of the first antibody includes HCDR1 (SEQ ID NO: 11), HCDR2 (SEQ ID NO: 12), and HCDR3 (SEQ ID NO: 13), and the light chain variable region of the first antibody includes LCDR1 (SEQ ID NO: 15), LCDR2 (SEQ ID NO: 16), and LCDR3 (SEQ ID NO: 17); and 2) A second antibody conjugated with acridinium ester, wherein the heavy chain variable region of the second antibody includes HCDR1 with sequence SEQ ID NO:3, HCDR2 with sequence SEQ ID NO:4, and HCDR3 with sequence SEQ ID NO:5, and the light chain variable region of the first antibody includes LCDR1 with sequence SEQ ID NO:7, LCDR2 with sequence SEQ ID NO:8, and LCDR3 with sequence SEQ ID NO:9.
The myosin-binding protein C detection kit as described in claim 1, wherein: 1) The heavy chain variable region sequence of the first antibody is shown in SEQ ID NO:2, and the light chain variable region sequence of the first antibody is shown in SEQ ID NO:6; the heavy chain variable region sequence of the second antibody is shown in SEQ ID NO:10, and the light chain variable region sequence of the second antibody is shown in SEQ ID NO:14; or 2) The heavy chain variable region sequence of the first antibody is shown in SEQ ID NO:10, and the light chain variable region sequence of the first antibody is shown in SEQ ID NO:14; the heavy chain variable region sequence of the second antibody is shown in SEQ ID NO:2, and the light chain variable region sequence of the first antibody is shown in SEQ ID NO:6.
The myosin-binding protein C detection kit as described in claim 1 further includes myosin-binding protein C standard solutions of different concentrations.
Use of the myosin-binding protein C detection kit according to any one of claims 1-3 in the preparation of a diagnostic kit for diagnosing acute myocardial infarction.

Description

Cardiac myosin-binding protein C detection kit and its application Technical Field This invention relates to a myosin-binding protein C detection kit, and more particularly to a detection kit that can detect the content of myosin-binding protein C in serum with high sensitivity using a double antibody sandwich method. Background Technology Acute myocardial infarction (AMI) is a critical cardiovascular event caused by unstable ischemic syndrome, characterized by high incidence, poor prognosis, and high mortality. However, for patients with AMI who do not present with typical chest pain and show no significant changes on electrocardiogram (ECG), accurate diagnosis cannot be made solely based on ECG, echocardiography, and cardiac MRI. Currently, the globally recognized "gold standard" for diagnosing AMI uses cardiac troponin (cTn) as a marker of myocardial injury. cTn begins to be released 4–6 hours after the onset of AHI/AMI, is detectable in the blood, peaks at 18–24 hours, and disappears after two weeks. High-sensitivity cTnT products appear 2.5 hours after the onset of myocardial injury, but have low specificity, with a clinical false positive rate as high as 30%-50%, easily leading to misdiagnosis. European cardiac guidelines explicitly state that high-sensitivity cTn should not be used as a diagnostic criterion within 3 hours. Cardiac myosin-binding protein C (cMyBP-C) is currently the most recently studied cardiac biomarker for acute myocardial infarction (AHI)/acute myocardial infarction (AMI) both domestically and internationally. It exhibits high specificity and can specifically reflect myocardial lesions. In the early stages of AMI, large amounts of cMyBP-C and its metabolites are released into the bloodstream. Its elevated levels can be detected in the circulatory system as early as 30 minutes after myocardial injury, making it effective for early detection of myocardial injury. Furthermore, compared to other biomarkers, cardiac myosin-binding protein C is the first to be cleared from the bloodstream, thus possessing good prognostic value. For the detection of cardiac myosin-binding protein C (cMyBP-C), existing methods such as some myocardial injury quadruple test cards (colloidal gold method) and enzyme-linked immunosorbent assay (ELISA) kits are limited by methodological constraints. Their sensitivity is insufficient, and they cannot achieve quantitative detection. They cannot accurately assess the trend and risk of the disease. Furthermore, biomarkers are nonspecific, often resulting in false positives. Patients with renal insufficiency, pulmonary embolism, or stroke but without myocardial injury may have elevated biomarker indicators, leading to various misdiagnoses, delays, or overtreatment. As a novel biomarker, human cardiac myosin-binding protein C (cMyBP-C, MYBPC3, cMyC) currently lacks a method for accurate quantitative detection. Currently, the only authorized patent related to this biomarker detection is the cMyBP-C detection kit using a colloidal gold nanocage method developed by Nanjing Botiankezhi Biotechnology Co., Ltd. However, the colloidal gold method relies on the large aggregation of antibody-antigen complexes on the detection line, thus limiting its application to qualitative or semi-quantitative determination, often resulting in significant errors. Furthermore, the colloidal gold method exhibits a significant hook effect, restricting the linear range of the kit and hindering its clinical application in differentiating between weakly and strongly positive cases. Generally, current human cardiac myosin-binding protein C products only provide qualitative results with a reference cutoff value (positive threshold) of 50 pg/ml, which limits the clinical application of this biomarker in grading early myocardial injury and assessing the prognosis of cardiovascular surgeries. Summary of the Invention On one hand, this article provides a myosin-binding protein C detection kit, which includes: 1) Magnetic beads conjugated with a first antibody, wherein the heavy chain variable region of the first antibody includes HCDR1 (SEQ ID NO:3), HCDR2 (SEQ ID NO:4), and HCDR3 (SEQ ID NO:5), and the light chain variable region of the first antibody includes LCDR1 (SEQ ID NO:7), LCDR2 (SEQ ID NO:8), and LCDR3 (SEQ ID NO:9); and 2) A second antibody conjugated with acridinium ester, wherein the heavy chain variable region of the second antibody includes HCDR1 (SEQ ID NO: 11), HCDR2 (SEQ ID NO: 12), and HCDR3 (SEQ ID NO: 13), and the light chain variable region of the first antibody includes LCDR1 (SEQ ID NO: 15), LCDR2 (SEQ ID NO: 16), and LCDR3 (SEQ ID NO: 17). or 1) Magnetic beads conjugated with a first antibody, wherein the heavy chain variable region of the first antibody includes HCDR1 (SEQ ID NO: 11), HCDR2 (SEQ ID NO: 12), and HCDR3 (SEQ ID NO: 13), and the light chain variable region of the first antibody includes LCDR1 (SEQ ID NO: 15), LCDR2 (SEQ ID NO: 16), and LCDR3 (SEQ ID NO: 17); and