WO2023098135A1 - Kit de dosage immunologique à chimiluminescence photo-initiée contenant des microsphères luminescentes magnétiques, et son utilisation - Google Patents

Kit de dosage immunologique à chimiluminescence photo-initiée contenant des microsphères luminescentes magnétiques, et son utilisation Download PDF

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WO2023098135A1
WO2023098135A1 PCT/CN2022/111181 CN2022111181W WO2023098135A1 WO 2023098135 A1 WO2023098135 A1 WO 2023098135A1 CN 2022111181 W CN2022111181 W CN 2022111181W WO 2023098135 A1 WO2023098135 A1 WO 2023098135A1
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magnetic
microspheres
minutes
reaction system
luminescent
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PCT/CN2022/111181
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Chinese (zh)
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严义勇
朱海
马红圳
王嘉欣
邓炀
吴莹莹
梁健欣
钟锦威
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深圳市易瑞生物技术股份有限公司
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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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
    • G01N33/54326—Magnetic particles
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/76—Chemiluminescence; Bioluminescence

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  • the invention relates to the field of photochemiluminescence immunoassay, in particular to a photochemiluminescence immunoassay kit containing magnetic luminescent microspheres and an application thereof.
  • Photochemiluminescence immunoassay is a typical homogeneous immunoassay method. It is characterized by "double spheres", that is, “luminescent microspheres” and “photosensitive microspheres”, based on the antigen or antibody coated on the surface of these two microspheres, which is coupled with the target in the liquid phase to form an immune complex. Bring the two microspheres closer together. Under the action of excitation light, the "photosensitive microspheres” with photosensitive function can convert oxygen molecules in the surrounding environment into singlet oxygen, and transfer them to the "luminescent microspheres” with luminescent function, thereby inducing the formation of oxygen molecules on the luminescent microspheres. Partial chemiluminescent reaction produces high-energy red light.
  • the number of photons is converted to the target concentration by single photon counter and mathematical fitting.
  • the two types of microspheres cannot form immune complexes, and the distance between them will exceed the transmission range of singlet oxygen, and no high-energy red light signal will be generated.
  • this homogeneous immunoassay method directly measures the sample to be tested and the relevant reagents in the reaction system after mixing and reacting during the determination process, without redundant separation or cleaning steps, so it has rapid, separation-free and Cleaning, high sensitivity and easy operation are featured.
  • the sensitivity of this homogeneous immunoassay method may not meet the requirements due to the low concentration of microsphere complexes (or called microclusters) caused by the low concentration of target substances.
  • the purpose of the present invention is to provide an improved photochemiluminescent immunoassay method.
  • the present invention provides a kit containing magnetic luminescent microspheres and its application for photo-activated chemiluminescence immunoassay.
  • the inventors of the present application found that adding magnetic cores to the "luminescent microspheres" in the double-spheres of light-induced chemiluminescence immunoassay makes the luminescent microspheres magnetic, and the movement of the luminescent microspheres can be controlled by magnetism during the reaction process , so that the luminescent microspheres that capture the target can be separated from the reaction system and cleaned before the luminescent microspheres and photosensitive microspheres form a microsphere complex, thereby reducing background interference in subsequent detection, improving luminescent signal, and improving detection
  • the flexibility and adjustability of the method can realize various possibilities in methodology.
  • kits for immunodetection of a target by photoactivated chemiluminescence comprising:
  • the magnetic luminescent microspheres are labeled with the first antibody
  • Non-magnetic photosensitive microspheres labeled with a second binding moiety capable of binding to the first binding moiety.
  • a method for immunodetection of a target by photoactivated chemiluminescence which uses the kit described in the first aspect, and the method includes the following steps:
  • a method for immunodetection of a target by photoactivated chemiluminescence which uses the kit according to the first aspect, the method comprising the following steps:
  • the beneficial effects of the present invention are as follows: the luminescent microspheres (luminescent microsphere-target complexes) that capture the target on the antibody can be separated from the reaction system by using magnetism, and the complex is purified by the cleaning process, thereby reducing subsequent detection. Background interference in the medium; in the detection that requires high sensitivity, the complex can be transferred to a smaller reaction system using magnetism, thereby increasing the concentration of the complex, and then making singlet oxygen between the photosensitive microspheres and the luminescent microspheres The transmission can be more efficient, thereby improving the detection signal; the complex can be transferred in different reaction systems by using magnetism, so as to realize the process of binding and detection in different systems, and improve the flexibility and adjustability of detection.
  • Figure 1 shows the particle size distribution of magnetoluminescent microspheres prepared according to one embodiment of the present invention.
  • Fig. 2 shows the linear correlation curve between the low concentration range of 0 pg/ml to 150 pg/ml and the luminescence amount of gastrin releasing peptide precursor obtained by using non-magnetic luminescent microspheres.
  • Figure 3 shows the linear correlation curve between the full concentration range of the gastrin releasing peptide precursor from 0 pg/ml to 8000 pg/ml and the luminescence amount obtained by using non-magnetic luminescent microspheres.
  • Fig. 4 shows the linear correlation curve between the low concentration range of 0 pg/ml to 150 pg/ml and the luminescence amount of the gastrin releasing peptide precursor obtained by using the magnetic luminescent microspheres of the present invention.
  • Fig. 5 shows the linear correlation curve of the gastrin releasing peptide precursor in the full concentration range from 0 pg/ml to 8000 pg/ml and the luminescence amount obtained by using the magnetic luminescent microspheres of the present invention.
  • Fig. 6 shows the linear correlation curve between the concentration of procalcitonin in the range of 0 ng/ml to 30 ng/ml and the amount of luminescence obtained by using non-magnetic luminescent microspheres.
  • Fig. 7 shows the linear correlation curve between the concentration range of procalcitonin from 0 ng/ml to 30 ng/ml and the amount of luminescence obtained by using magnetic luminescent microspheres.
  • the present application provides a kit for immunodetection of a target by photoactivated chemiluminescence, which includes:
  • the magnetic luminescent microspheres are labeled with the first antibody
  • Non-magnetic photosensitive microspheres labeled with a second binding moiety capable of binding to the first binding moiety.
  • the first binding moiety and the second binding moiety are selected from a pair of substances capable of specifically binding to each other, such as ligands, oligonucleotides, oligonucleotide binding proteins, Lectin, hapten, antigen, immunoglobulin binding protein, avidin, avidin or biotin.
  • the first binding moiety is one of avidin and biotin
  • the second binding moiety is the other of avidin and biotin.
  • the avidin may be, for example, avidin, vitellavidin, streptavidin, neutravidin, or avidin-like, but not limited thereto.
  • the first binding moiety is one of streptavidin and biotin
  • the second binding moiety is the other of streptavidin and biotin.
  • said first binding moiety is biotin and said second binding moiety is streptavidin.
  • the combination between the first binding part and the second binding part can make the second antibody bind to the non-magnetic photosensitive microsphere, and the second antibody can bind to the magnetic luminescent microsphere.
  • the first antibody bound to the target forms a double-antibody sandwich structure, thereby shortening the distance between the magnetic luminescent microspheres and the non-magnetic photosensitive microspheres, so that the chemiluminescent reaction can occur under the condition of light excitation. Therefore, those skilled in the art can also select appropriate first binding moieties and second binding moieties to label the second antibody and non-magnetic photosensitive microspheres respectively according to needs.
  • binding in the context of the present invention has a broad meaning as understood by those skilled in the art, and specifically refers to binding due to, for example, covalent coupling, coordination, electrostatic, hydrophobic, ionic and/or hydrogen bonding A direct association between two molecules caused by an equal interaction.
  • the target can be a disease-related marker, for example, a tumor marker, such as gastrin releasing peptide precursor, alpha-fetoprotein, carbohydrate antigen, etc.; an inflammatory disease marker, Such as procalcitonin, interleukin, C-reactive protein, etc.; virus-associated antigens, such as African swine fever, bovine foot-and-mouth disease, bovine viral diarrhea virus, etc.
  • a tumor marker such as gastrin releasing peptide precursor, alpha-fetoprotein, carbohydrate antigen, etc.
  • an inflammatory disease marker such as procalcitonin, interleukin, C-reactive protein, etc.
  • virus-associated antigens such as African swine fever, bovine foot-and-mouth disease, bovine viral diarrhea virus, etc.
  • the target substance can also be a drug and its metabolites, such as antibacterial drugs, antifungal drugs, antiviral drugs, antitumor agents, steroids, hormones, etc. for humans or animals and its metabolites.
  • a drug and its metabolites such as antibacterial drugs, antifungal drugs, antiviral drugs, antitumor agents, steroids, hormones, etc. for humans or animals and its metabolites.
  • the magnetic luminescent microspheres are prepared by the following method:
  • the luminescent composition contains an olefin compound and a metal chelate
  • the Fe 3 O 4 magnetic beads are prepared from ferric chloride or its hydrate.
  • the magnetic luminescent microspheres use Fe3O4 magnetic beads as the core, and the surface is coated with polymers with active groups such as carboxyl groups, amino groups, aldehyde groups, epoxy groups, azo groups, olefins, and alkynes, such as Polystyrene, polycaprolactone, agarose, silica, etc. This reactive group can be used to conjugate antibodies.
  • active groups such as carboxyl groups, amino groups, aldehyde groups, epoxy groups, azo groups, olefins, and alkynes, such as Polystyrene, polycaprolactone, agarose, silica, etc. This reactive group can be used to conjugate antibodies.
  • said luminescent composition comprises an alkene compound and a metal chelate.
  • the olefinic compound may be 2-phenyloxathiene and its derivatives.
  • the metal of the metal chelate may be a fluorescent rare earth metal, preferably selected from yttrium, europium, gadolinium, lanthanum, cerium, terbium, ytterbium, samarium, etc., more preferably europium.
  • the metal chelate is a europium (Eu) complex, such as (1,10-phenanthroline) tris[4,4,4-trifluoro-1-(2-thienyl)-1,3 - butanedione] europium(III).
  • Eu europium
  • the particle size of the magnetic luminescent microspheres is 40nm to 800nm. In a further preferred embodiment, the particle size of the magnetic luminescent microspheres is 100 nm to 300 nm.
  • the particle size of the non-magnetic photosensitive microspheres may be 40 nm to 800 nm. In a preferred embodiment, the non-magnetic photosensitive microspheres have a particle diameter of 100 nm to 300 nm.
  • Non-magnetic photosensitive particles are polymer particles filled with photosensitive compounds.
  • the photosensitive compound may be, for example, a phthalocyanine dye, a porphyrin derivative, or other compounds that can receive light and generate active oxygen.
  • the non-magnetic photosensitive microparticles can be obtained commercially, for example, from PerkinElmer Co., Ltd. Those skilled in the art can select non-magnetic photosensitive particles suitable for the present invention according to actual needs. Before being used in the present invention, those skilled in the art can use conventional means in the art to label the second binding moiety on commercially available non-magnetic photosensitive microspheres.
  • a method for immunodetection of a target by photoactivated chemiluminescence which uses the kit according to the first aspect, the method comprising the following steps:
  • sample to be tested refers to a sample to be tested that contains or is suspected to contain the target substance to be tested.
  • Samples to be tested that can be used in the present invention include body fluids, such as human or animal serum, plasma, urine, sputum, milk, saliva; solvents; food samples such as vegetables and fruits; environmental samples such as soil or water samples; plant materials; Cells; Bacteria; Viruses; Fungi, etc.
  • the concentration of the magnetic luminescent microspheres in the first reaction system or the second reaction system is 0.001 mg/mL to 5 mg/mL;
  • the concentration in the system or the second reaction system is 0.001 mg/mL to 5 mg/mL;
  • the concentration of the non-magnetic photosensitive microspheres in the second reaction system is 0.001 mg/mL to 5 mg/mL.
  • the concentration of the magnetic luminescent microspheres in the first reaction system or the second reaction system is 0.04 mg/mL to 0.4 mg/mL;
  • the concentration in the reaction system or the second reaction system is 0.04 mg/mL to 0.4 mg/mL; and the concentration of the non-magnetic photosensitive microspheres in the second reaction system is 0.04 mg/mL to 0.4 mg/mL.
  • step a) comprises incubating at a temperature ranging from 35°C to 40°C for 5 minutes to 20 minutes, for example at 37°C for 15 minutes.
  • the magnetic separation in step b) may include using tools well known in the art such as a magnetic rod, a magnetic plate, etc., without any special limitation.
  • the separated magnetic luminescent microspheres are transferred to a washing solution for washing.
  • a washing solution for washing.
  • a cleaning solution such as HEPES buffer, PBS buffer, TRIS-HCl and the like. Washing can separate the target captured by the first antibody on the magnetic luminescent microspheres from the first reaction system, and the magnetic luminescent microsphere-target complex can be purified by the washing process to reduce background interference.
  • step c) comprises incubating at a temperature ranging from 35°C to 40°C for 5 minutes to 15 minutes, for example at 37°C for 15 minutes.
  • step d) comprises incubating at a temperature ranging from 35°C to 40°C for 5 minutes to 15 minutes, for example at 37°C for 15 minutes.
  • the magnetic luminescent microspheres labeled with the first antibody will be coupled to the target object through the first antibody; after the incubation in step c), the second The second antibody is coupled to the target; after the incubation in step d), the second antibody and the non-magnetic photosensitive microspheres are combined through the first binding part and the second binding part; and, the first antibody and the second Different binding sites of antibodies on the target are coupled to the target.
  • said first reaction system and said second reaction system are the same or different.
  • the microsphere complex after magnetic separation can be transferred to a system different from the first reaction system, so that the binding and detection process between microspheres, target objects and antibodies can be carried out in different systems, thereby improving the flexibility of detection and adjustability.
  • the second reaction system may have a different solvent, pH, surface activity concentration, etc. from the first reaction system, so that the combination of the second antibody or the photosensitive microsphere and the microsphere complex can be promoted, and the detection efficiency can be reduced. background noise etc.
  • solution volume of the first reaction system and the solution volume of the second reaction system may be the same or different.
  • the microsphere complexes after magnetic separation can be transferred to the second reaction system with a smaller volume, thereby increasing the concentration of the microsphere complexes, making the microsphere complexes closer to each other, and making The transfer of reactive oxygen species between photosensitive microspheres and luminescent microspheres is more efficient, thereby improving the detection signal. Therefore, in a preferred embodiment, the solution volume of the second reaction system is smaller than the solution volume of the first reaction system.
  • methods well known in the art can be used to detect the luminous intensity in step e), for example, firstly, laser light is used, such as using 600nm to 700nm light to excite non-magnetic photosensitive particles, and oxygen in the air can be Molecules are converted into singlet oxygen. When the distance between the non-magnetic photosensitive particles and the magnetic luminescent particles is close enough, the singlet oxygen can be transferred to the magnetic luminescent particles, react with the luminescent compounds in the luminescent particles, and excite the metal chelate in them. metals, eventually producing short-wavelength photons such as 520nm to 620nm. Then, a commercial microplate reader can be used to detect the luminescence intensity of the magnetic luminescent microspheres. The detected luminescence intensity determines whether the target substance is contained in the sample to be tested and the content of the target substance.
  • a method for immunodetection of a target by photoactivated chemiluminescence which uses the kit according to the first aspect, the method comprising the following steps:
  • the concentration of the magnetic luminescent microspheres in the first reaction system or the second reaction system is 0.001 mg/mL to 5 mg/mL;
  • the concentration in the system or the second reaction system is 0.001 mg/mL to 5 mg/mL;
  • the concentration of the non-magnetic photosensitive microspheres in the second reaction system is 0.001 mg/mL to 5 mg/mL.
  • the concentration of the magnetic luminescent microspheres in the first reaction system or the second reaction system is 0.04 mg/mL to 0.4 mg/mL;
  • the concentration in the reaction system or the second reaction system is 0.04 mg/mL to 0.4 mg/mL, and the concentration of the non-magnetic photosensitive microspheres in the second reaction system is 0.04 mg/mL to 0.4 mg/mL.
  • step a) comprises incubating at a temperature ranging from 35°C to 40°C for 5 minutes to 20 minutes, for example at 37°C for 15 minutes.
  • the magnetic separation in step b) may include using tools known in the art such as a magnetic rod, a magnetic plate, etc., and there is no special limitation.
  • the separated magnetic luminescent microspheres are transferred to a washing solution for washing.
  • a washing solution for washing.
  • a cleaning solution such as HEPES buffer, PBS buffer, TRIS-HCl and the like. Washing can separate the target captured by the first antibody on the magnetic luminescent microspheres from the first reaction system, and the magnetic luminescent microsphere-target complex can be purified by the washing process to reduce background interference.
  • step c) comprises incubating at a temperature in the range of 35°C to 40°C for 5 minutes to 15 minutes, for example at 37°C for 15 minutes.
  • the magnetic luminescent microspheres labeled with the first antibody will be coupled to the target through the first antibody, and the second antibody will also be coupled to the target. target object coupling; and, the different binding sites of the first antibody and the second antibody on the target object are coupled with the target object; after step c) incubation, the second antibody and the non-magnetic photosensitive microsphere Binding is via said first binding moiety and second binding moiety.
  • the inventors of the present application found that whether the photosensitive microsphere-target complex is separated from the system before coupling with the second antibody and then coupled with the second antibody, or the second antibody is coupled to the photosensitive microsphere- After the target complex forms a microsphere complex and is separated from the system, the technical effects of reducing background interference, improving detection signal, and improving detection flexibility and adjustability can be achieved.
  • the separation is performed after the second antibody is coupled to the photosensitive microsphere-target complex to form a microsphere complex, which is useful for reducing background interference, improving detection signal, and improving detection flexibility and reliability. Tonality is better.
  • said first reaction system and said second reaction system are the same or different.
  • the microsphere complex after magnetic separation can be transferred to a system different from the first reaction system, so that the binding and detection process between microspheres, target objects and antibodies can be carried out in different systems, thereby improving the flexibility of detection and adjustability.
  • the second reaction system may have a different solvent, pH, surface activity concentration, etc. from the first reaction system, so that the combination of the second antibody or the photosensitive microsphere and the microsphere complex can be promoted, and the detection efficiency can be reduced. background noise etc.
  • solution volume of the first reaction system and the solution volume of the second reaction system may be the same or different.
  • Fe 3 O 4 magnetic core Add 16.2g of ferric chloride hexahydrate, 500ml of polyethylene glycol and 1g of sodium acetate into a 1000ml three-necked flask, vacuumize and nitrogen, 300rpm magnetic stirring, reflux at 200°C for 48 hours, ultrapure water Wash three times to get the magnetic core.
  • Magnetic polymer microspheres Get 1g of the magnetic core obtained above and add 30ml absolute ethanol, add 0.5ml methacrylic acid, 0.25mL polystyrene (average molecular weight 26000), import into 250ml 75 ethanol with 450rpm mechanical Add 5 ml of ethanol and 0.1 mM azobisisobutyronitrile into the stirred three-necked flask, blow nitrogen to remove oxygen, heat to 75° C., and stir overnight to obtain magnetic polymer microspheres.
  • Magnetic luminescent microspheres Take 60 mg of the above-mentioned magnetic polymer microspheres, set the volume to 30 mL, vortex-coat to prepare luminescent microspheres, add 2.85 mL of dichloroethane, add Eu complex (1,10-phenanthroline ) Tris[4,4,4-trifluoro-1-(2-thienyl)-1,3-butanedione]europium(III) 20mg, 1-xylylenediamino, 2-phenyloxathia 10 mg of cyclohexene was vortexed for 2 hours, then evaporated in a rotary evaporator at 40° C. for 30 minutes, and then washed 3 times with absolute ethanol to obtain magnetic luminescent microspheres.
  • Particle size characterization Take 10 ⁇ L of the magnetic luminescent microspheres prepared in Example 1, disperse them in ultrapure water, control the concentration in the range of 0.1-1 mg/mL, and use a laser particle size analyzer to test, the results are shown in Figure 1. It can be seen that the average particle diameter of the magnetic luminescent microspheres prepared according to Example 1 of the present invention is 182 nm, and the PDI is 0.044.
  • Characterization of carboxyl group content Take 100 mg of the magnetic luminescent microspheres prepared in Example 1, disperse them in 100 mL of ultrapure water, and use 0.1 M sodium hydroxide solution to titrate with a potentiometric titrator, and calculate the carboxyl group content to be 86 ⁇ mol/g. In terms of experimental operability, magnetic luminescent microspheres with a carboxyl content in the range of 60 ⁇ mol/g to 200 ⁇ mol/g are usually selected.
  • Example 3 Labeling Magnetic Luminescent Microspheres with Primary Antibody
  • Activation Take 100 ⁇ L of 10 mg/mL magnetic luminescent microspheres (ie 1 mg), centrifuge at 10,000 rpm for 15 minutes, remove the supernatant, add 200 ⁇ L of pH7.0 100mM HEPES buffer, and resuspend by ultrasonication.
  • N-hydroxysuccinimide NHS
  • EDC 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride
  • Blocking After the coupling step, add 100 ⁇ L blocking agent, then block overnight at 37°C; replace with HEPES solution after overnight.
  • Example 4 Linear correlation between target concentration and luminous intensity
  • non-magnetic luminescent microspheres as a control, add 25 ⁇ L of non-magnetic luminescent microspheres labeled with antibody I of gastrin-releasing peptide precursor to the microtiter plate, and 10 ⁇ L of gastrin-releasing peptide precursor at a concentration ranging from 0 pg to 8000 pg Serum samples of different concentrations to be tested and 25 ⁇ L of biotinylated antibody II were incubated at 37°C for 15 minutes; then 75 ⁇ L of streptavidin-labeled photosensitive microspheres were added to it and incubated at 37°C for 15 minutes, and then the luminescence intensity was measured.
  • Example 5 Testing of particle size and luminous intensity of magnetic luminescent microspheres
  • Table 1 Luminescence intensity of samples with different concentrations (ng/mL) under magnetic luminescent microspheres with different particle sizes (nm)
  • the concentration of procalcitonin in the serum sample to be tested was compared with the luminescent results obtained by the magnetic luminescent microspheres and non-magnetic luminescent microspheres respectively, and the results are shown in Figures 6-7 and Table 2 below.
  • the signal difference when the concentration is 0 may be the influence of the serum matrix, which comes from non-specific adsorption in the matrix. It can be seen from Table 2 that this matrix effect makes it impossible for non-magnetic luminescent microspheres to distinguish the three concentrations of 0ng/mL, 0.008ng/mL and 0.04ng/mL, while magnetic luminescent microspheres avoid Influence of matrix effect. It can be seen that the detection limit using magnetic luminescent microspheres reduces the detection limit.
  • the luminous intensity value of using magnetic luminescent microspheres is at least 2 times that of non-magnetic luminescent microspheres, which is obviously more conducive to the distinction of concentration. That is to say, the detection using magnetic luminescent microspheres is beneficial to the distinction of concentration, with reduced detection limit and improved accuracy.
  • the concentration of the gastrin-releasing peptide precursor in the serum sample to be tested was compared with the luminescent results obtained by the magnetic luminescent microspheres and non-magnetic luminescent microspheres respectively, and the results are shown in Table 3 below. It can be seen from the data in the table that although the two concentrations of 0ng/mL and 0.0016ng/mL can be clearly distinguished by using non-magnetic luminescent microspheres and magnetic luminescent The luminescence intensity obtained by detecting the concentration of 0.0016ng/mL is 3 times that of 0ng/mL, while the luminescence intensity obtained by detecting the concentration of 0.0016ng/mL by using the ratio of non-magnetic luminescent particles is less than 2 times that of detecting the concentration of 0ng/mL. This shows from the side that the detection limit of detection using magnetic luminescent particles is significantly lower, which is more conducive to the distinction of concentration and the improvement of detection accuracy.
  • Embodiment 8 Change the detection of reaction system

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Abstract

La présente invention concerne un kit de dosage immunologique à chimiluminescence photo-initiée comprenant : des microsphères luminescentes magnétiques marquées avec des premiers anticorps ; des seconds anticorps marqués avec des premiers fragments de liaison et des épitopes cibles différents du même antigène ; des microsphères photosensibles non magnétiques marquées avec des seconds fragments de liaison, les seconds fragments de liaison pouvant se lier aux premiers fragments de liaison. La présente invention concerne également l'utilisation du kit pour effectuer la détection d'un objet cible.
PCT/CN2022/111181 2021-11-30 2022-08-09 Kit de dosage immunologique à chimiluminescence photo-initiée contenant des microsphères luminescentes magnétiques, et son utilisation WO2023098135A1 (fr)

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CN116429752A (zh) * 2023-02-13 2023-07-14 科美博阳诊断技术(上海)有限公司 Afp检测试剂盒及其使用方法
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CN116380883B (zh) * 2023-02-13 2024-02-23 上海索昕生物科技有限公司 用于光激化学发光检测的感光微球

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