WO2022077931A1 - Procédé et kit de détection de troponine i cardiaque - Google Patents

Procédé et kit de détection de troponine i cardiaque Download PDF

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Publication number
WO2022077931A1
WO2022077931A1 PCT/CN2021/097947 CN2021097947W WO2022077931A1 WO 2022077931 A1 WO2022077931 A1 WO 2022077931A1 CN 2021097947 W CN2021097947 W CN 2021097947W WO 2022077931 A1 WO2022077931 A1 WO 2022077931A1
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WIPO (PCT)
Prior art keywords
antibody
cardiac troponin
kit
antibodies
amino acid
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PCT/CN2021/097947
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English (en)
Chinese (zh)
Inventor
刘春艳
汪荣
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广东菲鹏生物有限公司
菲鹏生物股份有限公司
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Priority to KR1020237016651A priority Critical patent/KR20230110263A/ko
Publication of WO2022077931A1 publication Critical patent/WO2022077931A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6887—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids from muscle, cartilage or connective tissue
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
    • G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
    • G01N2333/4701—Details
    • G01N2333/4712—Muscle proteins, e.g. myosin, actin, protein
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/32—Cardiovascular disorders
    • G01N2800/324—Coronary artery diseases, e.g. angina pectoris, myocardial infarction

Definitions

  • the present application relates to the field of protein detection. Specifically, the present application relates to a detection method and kit for cardiac troponin I.
  • cTnI cardiac troponin I
  • Cardiac troponin exists in trace amounts in normal human body, but when myocardial infarction occurs, the damage of myocardial cells results in the release of cTnI into the blood. Regardless of ST segment elevation, cTnI rises sharply, peaking around 24 hours, and falling to normal levels after 7-10 days. The magnitude of cTnI elevation was related to the degree of myocardial cell necrosis to a certain extent. If the cTnI in the patient's blood rises again during the descending process, it means that the myocardium is damaged/necrotic again. Changes in cTnI concentrations are clearly related to the risk of ACS (Acute Coronary Syndrome), treatment decisions and prognosis.
  • ACS Acute Coronary Syndrome
  • cardiac troponin I After a lot of theoretical research and experimental exploration, the inventors have fully considered the structural properties of cardiac troponin I, analyzed and studied various troponin antigen ranges and detection antibodies to be tested, and through a large number of cross-experiments and screenings, obtained can be used. A combination of cardiac troponin I antigen regions for cardiac troponin I detection. The inventors have demonstrated that antibodies that bind specific antigenic fragments are unexpectedly able to combine with each other for highly sensitive and specific detection of cTnI.
  • the present application provides a cardiac troponin I detection kit comprising a first antibody and a second antibody for detecting cardiac troponin I, wherein the first antibody binds to the first antibody of cardiac troponin I Amino acid fragment at positions 83-93; secondary antibody binds to the cardiac troponin CTNI-CTNC complex.
  • the second antibody is a coated antibody when the first antibody is a labeled antibody; or the first antibody is a coated antibody when the second antibody is a labeled antibody.
  • the kit further comprises a third antibody that detects cardiac troponin I, the third antibody binding to amino acid fragment 41-49 of cardiac troponin I;
  • the antibodies are divided into a first set of antibodies and a second set of antibodies for pairing, wherein the first set of antibodies and the second set of antibodies are selected from the group consisting of:
  • the first group of antibodies includes a first antibody
  • the second group of antibodies includes a second antibody and a third antibody
  • the first group of antibodies includes the second antibody, and the second group of antibodies includes the first antibody and the third antibody;
  • the first set of antibodies includes the third antibody
  • the second set of antibodies includes the first antibody and the second antibody.
  • the reactivity of the antibody is OD405 > 0.5 as assessed by ELISA.
  • the labeled antibody is labeled with at least one of a detectable label and a binding partner.
  • Detectable labels can be, for example, metal particles, fluorescent labels, chromophore labels, electron dense labels, chemiluminescent labels, radioactive labels, or enzymatic labels.
  • the detectable label can be, for example, rhodamine, fluorescein, fluorescent microspheres, colloidal gold, acridine ester, luciferase, horseradish peroxidase, alkaline phosphatase, ⁇ -galactosidase, Labeled with glucoamylase, lysozyme, sugar oxidase, glucose oxidase, galactose oxidase or glucose-6-phosphate dehydrogenase.
  • the binding partner can be, for example, biotin/avidin or biotin/streptavidin.
  • the coated antibody is attached to a solid phase and/or a binding partner, which may be, for example, magnetic particles, latex particles, microtiter plates, nitrocellulose membranes, or microfluidic chips ; the binding partner can be, for example, biotin/avidin, biotin/streptavidin; for example, the coated antibody is attached to the solid phase through the binding partner.
  • a binding partner which may be, for example, magnetic particles, latex particles, microtiter plates, nitrocellulose membranes, or microfluidic chips ;
  • the binding partner can be, for example, biotin/avidin, biotin/streptavidin; for example, the coated antibody is attached to the solid phase through the binding partner.
  • the application provides an antibody combination for detecting cardiac troponin I, including a first antibody and a second antibody; optionally, also includes a third antibody; wherein the first antibody, the second antibody, and the third antibody are as above definition.
  • the present application provides the use of the antibody combination as described above in the preparation of a kit for detecting cardiac troponin I.
  • kits are used for 1) dynamic detection of cardiac troponin levels, eg, for real-time monitoring, and/or 2) diagnosis of early acute myocardial injury, including, eg, excluding or Prediction of early acute myocardial injury.
  • the application provides a method for preparing the antibody combination as described above, the method comprising:
  • the kit of the present application includes a first antibody and a second antibody for detecting cardiac troponin I, wherein the first antibody binds to positions 83-93 of cardiac troponin I Amino acid fragment; the secondary antibody binds to the cardiac troponin CTNI-CTNC complex; for example, when the primary antibody is a labeled antibody, the secondary antibody is a coating antibody; or when the secondary antibody is a labeled antibody, the primary antibody is a coating antibody, and can be Improve detection specificity.
  • the kit of the present application includes a cardiac troponin I detection reagent card (test strip).
  • immunofluorescence techniques can be used to label antibodies on detectable labels such as fluorescent microspheres, and a double antibody sandwich method can be used to detect cTnI in subjects such as patients with myocardial injury/necrosis.
  • the methods and kits of the present application can improve detection specificity and/or reduce missed detections.
  • the present application may provide methods for the detection of cardiac troponin I by fluorescent immunochromatography or provide reagents comprising performing fluorescent immunochromatography.
  • immunochromatography can be accomplished by a double-antibody sandwich method with test strips.
  • the reagents and devices for performing the double-antibody sandwich method are not particularly limited, and any suitable reagents and/or devices may be employed.
  • fluorescent immunochromatographic test strips can include sample pads, binding pads, nitrocellulose (NC) membranes, and absorbent pads.
  • at least one fluorophore-labeled antibody is immobilized on the binding pad.
  • the NC membrane can be coated with two lines, eg, a test (T) line and a control (C) line, respectively; the T line can be coated with at least one antibody and can capture the The fluorophore-labeled antibody on the pad forms a complex with the antigen in the sample, thereby forming a double-antibody sandwich.
  • the sample to be tested is added to the sample loading port of the detection reagent card, and under the action of the lateral capillary, the sample to be tested first passes through the binding pad, and the fluorophore-labeled antibody on the binding pad undergoes specific binding.
  • Immunobinding each binds to form an antigen-antibody fluorescent complex, which is immobilized in the T-line.
  • the C line is coated with a substance that reacts with the free fluorophore-labeled antibody.
  • the free fluorophore-labeled antibody passes through the C line, it can specifically immunocombine with the substance on the C line, thereby being immobilized on the C line. in line.
  • the fluorescence intensities of the C-line and T-line are contrasted, so that individual differences between samples can be corrected.
  • the fluorescence intensity of the two lines detected by the fluorescence immunoassay analyzer is reflected in the peak area, and the T/C value (T peak area/C peak area) is calculated by the instrument's own calculation software and fitted to the set standard curve.
  • the fluorescence immunoassay analyzer automatically converts the concentration value of cTnI in the corresponding sample to be tested.
  • the present application can use antibodies that bind different amino acid fragments to recognize multiple positions on the antigen, thereby reducing the risk of missed detection and improving the detection rate.
  • an antibody that binds to the 83-93aa fragment of cardiac troponin I is used as the labeling antibody, and an antibody that binds to the cardiac troponin I-C complex and 41-49aa that binds to cardiac troponin I is used Fragmented antibodies are mixed as coating antibodies.
  • an antibody that binds to the cardiac troponin I-C complex is used as the labeling antibody, and an antibody that binds the 83-93aa fragment of cardiac troponin I and 41-49aa that binds cardiac troponin I Fragmented antibodies are mixed as coating antibodies.
  • an antibody that binds to the 41-49aa fragment of cardiac troponin I is used as the labeling antibody, and an antibody that binds to the 83-93aa fragment of cardiac troponin I is complexed with an antibody that binds to the cardiac troponin I-C
  • the antibody of the substance was mixed as the coating antibody.
  • the present application provides a cardiac troponin I detection method, detection kit and preparation method.
  • the kit of the present application has improved sensitivity and specificity.
  • the antibodies of the present application may be monoclonal or polyclonal antibodies.
  • the antibodies of the present application can be prepared using methods known in the art.
  • the antibodies of the present application can be prepared by immunizing animals with antigens comprising the amino acid fragments described herein.
  • carrier proteins including but not limited to BSA, ovalbumin, KLH, etc.
  • immunoreactive substances eg, epitope peptides.
  • Carrier proteins can include proteins or polypeptides that can function as immunogenic carriers. These types of polypeptides include albumin, serum protein, globulin, crystallin, lipoprotein and/or fragments thereof.
  • immunoreactive substances eg, cardiac troponin I amino acid fragments, cardiac troponin CTNI-CTNC complex, but not limited thereto
  • binding of an antibody to an amino acid fragment of cardiac troponin may refer to the ability of the antibody to bind to the amino acid fragment, although the amino acid fragment is not necessarily the smallest binding fragment.
  • any suitable in vitro assay, cell-based assay, in vivo assay, animal model, etc. can be used to test the effects of the antibodies of the present application, such as binding activity and/or cross-reactivity.
  • the assay can include, for example, ELISA, FACS binding assay, Biacore, competitive binding assay, and the like.
  • the reactivity of the antibodies of the present application in binding to an antigen (antigenic peptide) is characterized, for example, in an ELISA, for example, as determined by reading a reaction value ⁇ 0.5 at 405 nm in a peroxidase-labeled ELISA method for better reactivity.
  • the kit includes the first antibody and the second antibody as described above; in one or more embodiments, the kit also includes the third antibody as described above; in one or more embodiments, other antibodies may also be used as coating or labeling antibodies.
  • the coated antibody is bound to a solid phase.
  • the coated antibody can be used to coat a solid support.
  • the solid support is not particularly limited, and it can be, for example, magnetic particles, latex particles, microtiter plates, nitrocellulose membranes, or microfluidic chips.
  • the coated antibody binds to a binding partner such as biotin/avidin, biotin/streptavidin; in one or more In embodiments, the coated antibody is attached to the solid phase via a binding partner.
  • the labeled antibody is labeled with a detectable label, such as a fluorescent label; in one or more embodiments, the labeled antibody is linked to a binding partner, the binding partner Examples are biotin/avidin, biotin/streptavidin; in some embodiments, the labeled antibody is labeled with a detectable label by a binding partner.
  • the term "antibody” is used herein in the broadest sense, which may include full-length monoclonal antibodies, polyclonal antibodies, bispecific or multispecific antibodies, chimeric antibodies, and Antigen-binding fragments of antibodies so long as they exhibit the desired biological activity, such as binding to cTnI antigen.
  • Antibody fragments include portions of full-length antibodies, preferably the antigen-binding or variable regions thereof. Examples of antibody fragments include Fab, Fab', F(ab') 2 , Fd, Fv, dAb, complementarity determining region (CDR) fragments, single chain antibodies (eg, scFv), diabodies or domain antibodies.
  • the detectable labels used in this application are not particularly limited.
  • the labels may include, but are not limited to, metal particles, fluorescent labels, chromophore labels, electron-dense labels, chemiluminescent labels, radiolabels, or enzymatic labels, eg, by enzymatic reactions or molecular interactions for indirect detection.
  • exemplary labels include, but are not limited to, can be rhodamine, fluorescein, fluorescent microspheres, colloidal gold, acridine esters, luciferase, horseradish peroxidase, alkaline phosphate Enzymes, beta-galactosidase, glucoamylase, lysozyme, sugar oxidase, glucose oxidase, galactose oxidase or glucose-6-phosphate dehydrogenase label, among others.
  • kits of the present application include reagents suitable for performing immunoassays.
  • the kits of the present application can be used to perform immunoassays, such as ELISA, indirect immunofluorescence assay (IFA), radioimmunoassay (RIA), and other non-enzyme-linked antibody binding assays or methods.
  • immunoassays such as ELISA, indirect immunofluorescence assay (IFA), radioimmunoassay (RIA), and other non-enzyme-linked antibody binding assays or methods.
  • a sample of cTnI to be examined may include biological tissues, cells or body fluids in healthy or pathological states, eg blood samples, eg plasma, serum, blood products, eg semen or vaginal secretions.
  • the present application provides antibody combinations for the detection of cardiac troponin I, and their use in the manufacture of kits for the detection of cardiac troponin I.
  • the application provides combinations of immunoreactive polypeptides comprising amino acid fragments described herein and cardiac troponin CTNI-CTNC complexes, and their use in the preparation of antibodies for the detection of cardiac troponin I the use of.
  • the present application provides a method of making an antibody that detects cardiac troponin I, the method comprising adding an immunoreactive polypeptide comprising an amino acid fragment described herein and the CTNI-CTNC complex as Animals are immunized with the antigen or hapten, respectively, to prepare antibodies such as monoclonal or polyclonal antibodies that detect cardiac troponin I.
  • Monoclonal or polyclonal antibodies can be prepared by methods known in the art.
  • the application provides an immunoreactive polypeptide comprising an amino acid fragment described herein and the use of the CTNI-CTNC complex in the manufacture of a reagent or kit for the detection of cardiac troponin I.
  • the present application provides a cardiac troponin (cTnI) detection method and kit, using an antibody that binds to the 83-93aa fragment of cardiac troponin I and an antibody that binds to the cardiac troponin CTNI-CTNC complex to improve detection specificity , and further reduce the risk of missed detection by using an antibody that binds to the 41-49aa fragment of cardiac troponin I.
  • cTnI cardiac troponin
  • Immune animals Take 8-12-week-old BALB/c mice of the same strain as myeloma cells, mix well with 100 ⁇ g/mouse cTnI antigen and an equal amount of Freund's complete adjuvant, and inject them into the abdominal cavity of mice. Every 2 weeks, 100 ⁇ g/mouse cTnI antigen was thoroughly mixed with the same amount of incomplete Freund's adjuvant, and injected into the mouse intraperitoneally for multiple times for boosting immunization. After testing mouse serum (indirect ELISA method), those with a titer above 1:2000 can be used for fusion. Three days before fusion, the mice were boosted by intraperitoneal immunization again at a dose of 50 ⁇ g/mouse.
  • BALB/c mouse peritoneal macrophages were used as feeder cells.
  • BALB/c mice were sacrificed by pulling their necks, immersed in 75% alcohol, and in a clean bench, the abdominal skin was cut open with scissors under aseptic operation, the peritoneum was exposed, and 5 mL of RPMI 1640 basal medium was injected into the abdominal cavity with a syringe.
  • RPMI 1640 screening medium complete RPMI 1640 medium containing HAT
  • adjust the cell concentration to 1 ⁇ 10 5 cells/mL
  • mice Three days after the last immunization of the mice, the spleen was taken out under sterile conditions, placed in a petri dish, washed once with RPMI 1640 basal medium, and placed on a nylon mesh in a small beaker to grind and filter to prepare a cell suspension. Centrifuge, discard the supernatant, resuspend in RPMI 1640 basal medium, repeat this three times, and count.
  • Immune mouse spleen cells and mouse myeloma cells after PEG treatment, form a mixture of various cellular components, including unfused myeloma cells and immune spleen cells, myeloma cell synkaryon and immune spleen cells , and heterokaryons of myeloma cells and immune splenocytes. Only the latter can form hybridoma cells. For this purpose, unfused cells and homofused synkaryons must be removed from this multi-cell mixture and true hybrid cells selected. Therefore, on the 1st, 3rd, 5th, and 7th days after cell fusion, the culture medium was exchanged with the aforementioned HAT medium.
  • each culture well was aspirated, and the culture wells containing cTnI antibody in the culture medium were detected by indirect ELISA method.
  • Hybridoma cells were cloned by limiting dilution. After culturing, a single cell can be proliferated into a homologous cell clone; a cell line with better reactivity and stable secretion of anti-cTnI monoclonal antibody is obtained through reactivity screening.
  • Microtiter plates (Nunc, Maxisorb) were coated with 100 ⁇ L/well of coating buffer containing 2.5 ⁇ g/mL human cTnI (as antigen) for 1 hour at room temperature. Post-coating treatments were incubated in PBS buffer and 1% monoclonal antibody for 30 minutes. Then, wash with wash buffer. 100 ⁇ L/well of antibody samples were incubated for 1 hour at room temperature. Then wash 2 more times with washing solution. It was then incubated with 100 ⁇ L/well of detection antibody peroxidase-conjugated goat anti-mouse IgG diluted 1:40000 in PBS buffer for 1 hour at room temperature.
  • the peroxidase activity is measured by conventional methods (eg, the reaction value at 405 nm is read with an ELISA plate reader), and an antibody with OD 405 ⁇ 0.5 (with good reactivity) is obtained.
  • the IC antibody is the antibody of the CTNI-CTNC complex, and its preparation method is referred to above.
  • the CTNI-CTNC complex is replaced by the cTnI antigen, and the IC antibody is obtained by immunization, wherein the CTNI-CTNC complex can be purchased from Haipeptide Biology.
  • Antibodies with different binding fragments were selected for coating and labeling, and cross-pairing experiments were performed.
  • the screening process was as follows:
  • Blocking Add 100uL of blocking agent, mix well in the dark, and react at 37°C for 1h;
  • Quality control substance cTnI antigen, diluted to 50, 20, 5, 1.25ng/mL with PBS;
  • T/C represents the ratio of the T peak area to the C peak area, and the activity of the reaction pairing. The higher the T/C in the quality control and positive samples, the higher the activity.
  • the quality control concentration is 50ng/mL
  • the T/C value is less than 0.1 and no gradient is recorded as basically inactive.
  • the present application also uses the above-mentioned antibody preparation scheme, using cTnI amino acid fragment 83-93 coupled to carrier protein as an immunogen to prepare an antibody that binds to the 83-93aa fragment, for example, an OD 405 ⁇ 0.5 is evaluated by ELISA after reactivity screening reaction
  • the pairing specificity of antibody I83A-9 and IC antibodies 15A-6 and 16C-3 are all higher than 99%; using cTnI amino acid fragment 41-49 coupled to carrier protein as immunogen to obtain antibodies that bind to 41-49aa fragment, for example
  • the reactive screening reaction was ELISA to evaluate the antibody I41D-2 with OD 405 ⁇ 0.5, and the missed detection rate of pairing with the former two did not exceed 1%.

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Abstract

L'invention concerne un procédé de détection de troponine I cardiaque et un kit, qui appartiennent au domaine de la détection de protéines. Le kit comprend un premier anticorps et un second anticorps pour détecter la troponine I cardiaque, les anticorps étant choisis parmi les anticorps suivants : un premier anticorps se liant à un fragment d'acide aminé à des positions 83 à 93 de la troponine I cardiaque; et un second anticorps, qui est un anticorps se liant au complexe de troponine cardiaque CTNI-CTNC.
PCT/CN2021/097947 2020-10-16 2021-06-02 Procédé et kit de détection de troponine i cardiaque WO2022077931A1 (fr)

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CN202011114995.6A CN114384249A (zh) 2020-10-16 2020-10-16 一种心肌肌钙蛋白i检测方法和试剂盒

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EP1473567A1 (fr) * 2003-04-30 2004-11-03 Susann Eriksson Dosage immunologique de la troponine I cardiaque
CN111308084A (zh) * 2019-12-30 2020-06-19 菲鹏生物股份有限公司 一种超敏心肌肌钙蛋白i检测方法和试剂盒

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CN109239361B (zh) * 2018-09-29 2022-02-08 迈克生物股份有限公司 一种心肌肌钙蛋白i的检测试剂盒及其制备方法
CN110836973A (zh) * 2019-09-09 2020-02-25 浙江博实生物科技有限公司 检测肌钙蛋白及复合物的双标记试剂盒及制备与检测方法

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US20030040024A1 (en) * 1996-06-25 2003-02-27 Katrukha Alex G. Method and kit for the diagnosis of troponin I
EP1473567A1 (fr) * 2003-04-30 2004-11-03 Susann Eriksson Dosage immunologique de la troponine I cardiaque
CN111308084A (zh) * 2019-12-30 2020-06-19 菲鹏生物股份有限公司 一种超敏心肌肌钙蛋白i检测方法和试剂盒

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