65 results about "Abzyme" patented technology

The present invention belongs to the field of biological detection technology, and specifically relates to an anti-monkey C-reactive protein monoclonal antibody, an enzyme-linked immunosorbent assay kit and its application. Named CRP-13, CRP-13 includes a heavy chain variable region and a light chain variable region; the heavy chain variable region has three complementary determining regions CDR-H1 to CDR-H3, and the amino acid sequences are shown in SEQ ID NO.6-SEQ ID NO.8; the light chain variable region has three complementary determining regions CDR-L1 to CDR-L3, and the amino acid sequences are shown in SEQ ID NO.9-SEQ ID NO.11. The monoclonal antibody of the present invention can specifically bind to monkey C-reactive protein with high specificity. The above monoclonal antibody can be used to screen a paired monoclonal antibody, and a monkey C-reactive protein enzyme-linked immunosorbent assay kit is obtained based on the combination of the monoclonal antibodies, which can detect the content of monkey C-reactive protein in monkey serum, plasma or tissue fluid.

The invention relates to a human lactoferrin indirect competitive enzyme-linked immunosorbent assay kit and application thereof. The kit comprises an elisa plate, an anti-human lactoferrin antibody, an elisa secondary antibody, a confining liquid, a color developing agent and a stop solution. A nitrocellulose membrane in Western-Blot is replaced by an elisa plate with non-specific adsorption protein, and along with the addition of a sample, an anti-human lactoferrin antibody and an enzyme-labeled secondary antibody, a lactoferrin-primary antibody-secondary antibody compound is formed. And adding a color developing agent to carry out color development reaction, wherein the absorbance value of the sample is positively correlated with the content of the lactoferrin contained in the sample. And calculating the content of human lactoferrin in the sample according to the absorbance value. The method for detecting the human lactoferrin in the fermentation liquor has the advantages of reality, economy, high efficiency and the like.

The invention relates to a hybridoma cell strain VAND8, an anti-vanillin monoclonal antibody generated by the hybridoma cell strain VAND8 and application, and belongs to the field of immunochemistry. The hybridoma cell strain VAND8 provided by the invention can be used for preparing a high-titer vanillin-resistant monoclonal antibody, and the titer measured by an enzyme-linked immunosorbent assay (ELISA) method can reach 3.2 * 10 < 5 >; the vanillin-resistant monoclonal antibody provided by the invention is high in sensitivity and good in specificity, the 50% inhibition concentration IC50 on vanillin is 22.1 mu g / mL, and the cross reaction rate with vanillin structural analogues is low or even no cross reaction exists; the method can be used for measuring the content of vanillin.

The invention discloses an indirect ELISA (Enzyme-Linked Immunosorbent Assay) kit for rapidly detecting a porcine delta coronavirus IgA antibody, and relates to the field of immunological detection. The indirect ELISA kit comprises an ELISA plate coated with an antigen, the antigen is a recombinant PDCoV S2 protein, and the amino acid sequence of the antigen is as shown in SEQ ID NO. 2. According to the indirect ELISA kit, a recombinant PDCoV S2 protein is used as a coating antigen to capture a PDCoV specific IgA antibody, a goat anti-pig IgA antibody labeled by horse radish peroxidase is used as a detection antibody, a 'PDCoV-S2 protein-PDCoV-S2 specific IgA antibody-enzyme labeled antibody' trimer compound can be formed on an ELISA plate, and the PDCoV specific IgA antibody-enzyme labeled antibody can be used for detecting the PDCoV specific IgA antibody. And the result can be directly judged by measuring the color change of the substrate and the absorbance value, so that the detection efficiency can be obviously improved. The indirect ELISA kit can be used for clinical detection and immune evaluation of the PDCoV.

The present invention belongs to the field of biological detection technology, and specifically relates to a monkey high-mobility group protein B1 monoclonal antibody, an enzyme-linked immunosorbent assay kit and its application. Named HMGB1-11, it includes a heavy chain variable region and a light chain variable region; the heavy chain variable region has three complementary determining regions CDR-H1 to CDR-H3, and the amino acid sequences are shown in SEQ ID NO.6-SEQ ID NO.8; the light chain variable region has three complementary determining regions CDR-L1 to CDR-L3, and the amino acid sequences are shown in SEQ ID NO.9-SEQ ID NO.11. The monoclonal antibody of the present invention can specifically bind to monkey high-mobility group protein B1, and the monoclonal antibody can be paired with a monoclonal antibody after screening. Based on the monoclonal antibody combination, a monkey high-mobility group protein B1 enzyme-linked immunosorbent assay kit is constructed, which can detect the content of monkey high-mobility group protein B1 in monkey serum, plasma or tissue fluid.

The invention belongs to the technical field of biology, and particularly relates to a hybridoma cell strain BAP1C12, an anti-benzo [a] pyrene monoclonal antibody generated by the hybridoma cell strain BAP1C12 and application. The hybridoma cell strain BAP1C12 provided by the invention can be used for preparing a high-sensitivity anti-benzo [a] pyrene monoclonal antibody, and the titer measured by an enzyme-linked immunosorbent assay (ELISA) method can reach 2 * 10 < 5 >. The anti-benzo [a] pyrene monoclonal antibody provided by the invention is high in sensitivity and good in specificity, and the 50% inhibition concentration IC50 of the anti-benzo [a] pyrene monoclonal antibody to benzo [a] pyrene is 0.45 ng / mL. The anti-benzo [a] pyrene monoclonal antibody provided by the invention is high in sensitivity and good in specificity, the cross reactions of the anti-benzo [a] pyrene monoclonal antibody to the structural analogues of fluoranthene, benzo [b] fluoranthene, indeno [1, 2, 3-cd] pyrene, benzo [a] anthracene, benzo [k] fluoranthene, anthracene, phenanthrene, acenaphthene, fluorene and naphthalene are less than 3.5%, and the anti-benzo [a] pyrene monoclonal antibody can be applied to benzo [a] pyrene content determination.

The present application relates to the technical field of biotechnology, and particularly relates to a D-dimer chemiluminescence assay kit. The present application provides a D-dimer chemiluminescence assay kit, which comprises a magnetic particle suspension (immunomagnetic beads) coated with D-dimer antibody 1, alkaline phosphatase-labeled D-dimer antibody 2 (enzyme-labeled antibody) and a calibrant. The immunomagnetic beads and the enzyme-labeled antibody are prepared by chemical coupling and stored in a buffer containing a soluble high molecular material, which can effectively improve the detection sensitivity and repeatability. The kit can be used in combination with a substrate solution containing a chromogenic substrate such as adamantane and its derivative AMPPD or APS-5 for a full-automatic immune test system, and the content of D-dimer in plasma or whole blood can be determined by a chemiluminescence instrument. The kit has high detection sensitivity and good repeatability.

A method for producing an antibody κ-type light chain having enzyme activity or improved enzyme activity includes: a modification step for modifying a polynucleotide encoding an antibody κ-type light chain in which the variable region is a polypeptide comprising an amino acid sequence in which the 95th amino acid residue from the N-terminal of the variable region by the Kabat classification is a proline residue so that the proline residue is deleted or substituted to obtain a polynucleotide encoding an antibody κ-type light chain in which the variable region is a polypeptide comprising an amino acid sequence in which the 95th amino acid residue from the N-terminal of the variable region by the Kabat classification is deleted or an amino acid residue other than a proline residue; and an expression step for expressing the antibody κ-type light chain having enzyme activity by an intracellular or extracellular expression system using an expression vector including the polynucleotide encoding an antibody κ-type light chain from after the modification step.

The invention discloses a multiplex fluorescent quantitative PCR kit for identifying fusobacterium, which comprises reaction solutions A, each group of reaction solutions A comprises a specific primer, a specific probe, an internal standard and a buffer solution of at least one target bacterium; the reaction liquid B comprises a hot start Taq antibody enzyme, a UDG enzyme and dNTPs (deoxyribonucleoside triphosphates); the standard solution comprises host cell nucleic acid; the reference substance comprises plasmids of fragments of all target bacteria and host cell nucleic acid. The invention further provides a multiple fluorescent quantitative PCR method for identifying the fusobacterium, and the multiple fluorescent quantitative PCR kit for identifying the fusobacterium is adopted. According to the application, the screened specific genetic marker is utilized, and the highly specific primer is designed based on the specific genetic marker, so that the combination efficiency of the primer and the probe is improved, the cross reaction of the primer and the formation of a primer dimer are avoided, and the specific genetic marker is obtained through reasonable combination of different target bacteria. According to the invention, simultaneous detection of multiple targets can be realized.

The invention discloses a bridging antibody, an enzyme-labeled polymer as well as a preparation method and application of the bridging antibody. The bridging antibody is prepared by the following steps: performing enzyme digestion, purification, reduction, sulfydryl protection and coupling on a polyclonal antibody and an anti-DMT-biotin monoclonal antibody through pepsin, and then purifying to obtain a heterodimer product polyclonal antibody-Fab '-Linker-Fab'-anti-DMT-biotin; hRP is subjected to maleimide and then coupled with a sulfhydrylated polymer to obtain a polymer enzyme, and then the polymer enzyme is coupled with a small molecule DMT-biotin to obtain the enzyme-labeled polymer which is specifically combined with the bridging secondary antibody. The obtained bridging antibody and enzyme-labeled polymer are used for preparing a kit for three-stage amplification of immunohistochemical signals, through cooperation of all the components, low-abundance antigens in tissues can be better detected, and the false positive rate is greatly reduced.

The invention provides an anti-neocrown N protein human monoclonal antibody, an enzyme-linked immunosorbent assay kit and application of the anti-neocrown N protein human monoclonal antibody and the enzyme-linked immunosorbent assay kit. The anti-neocoronal N protein human monoclonal antibody is obtained from N protein of human immune neocoronal virus, a heavy chain and a light chain of the anti-neocoronal N protein human monoclonal antibody have variable regions and complementary determining regions, and the anti-neocoronal N protein human monoclonal antibody is specifically combined with neocoronal N protein; and through paired use of the antibodies, the kit has strong anti-interference capability, good specificity and high sensitivity on clinical new crown detection.

The invention belongs to the field of biotechnology products for plant virus detection, and particularly relates to a double-antibody sandwich enzyme-linked immunosorbent assay kit for grape berry necrosis viruses, a preparation method and a detection method.The double-antibody sandwich enzyme-linked immunosorbent assay kit comprises a microwell plate for ELISA, an enzyme-labeled antibody, a buffer solution, a developing solution and a stop buffer, the microwell plate is coated with a GINV specific polyclonal antibody, and the microwell plate is coated with a GINV specific polyclonal antibody. The enzyme-labeled antibody is a GINV specific polyclonal antibody labeled by horse radish peroxidase. The immunogen adopted by the invention is a GINV coat protein antigen expressed by genetic engineering. The invention provides a rapid, sensitive, simple and convenient method for detecting the grape berry necrosis virus on the serological level, is economical and practical, and also provides an effective method for preventing and treating the grape berry necrosis virus in grape production and preparing virus-free seedlings of the grape berry necrosis virus.

The invention discloses an electrochemiluminescence detection method of PARP enzyme, a kit and application thereof. The electrochemical luminescence detection method for detecting PARP enzyme comprises the following steps: (1) mixing human whole blood, a PARP inhibitor and a PARG inhibitor, incubating, and centrifuging to obtain human peripheral blood mononuclear cells; (2) collecting a human peripheral blood mononuclear cell precipitate for cracking, detecting the total protein concentration, adding a sample, adding a cell lysis solution into an electrochemical luminescent plate for incubation, and adding a PARP enzyme detection antibody for re-incubation; and (3) adding a detection secondary antibody for incubation to form a capture antibody-PARP enzyme-detection antibody-secondary antibody complex, and adding plate reading liquid for plate reading. The electrochemical luminescence detection method and the kit have the characteristics of high sensitivity, high specificity, high accuracy, high precision and simple operation method, and can quantitatively detect the content of PARP enzyme.

The present invention belongs to the field of biological detection technology, and specifically relates to an anti-monkey interleukin-6 monoclonal antibody, an enzyme-linked immunosorbent assay kit and its application. Named IL-6-12, IL-6-12 includes a heavy chain variable region and a light chain variable region; the heavy chain variable region has three complementary determining regions CDR-H1 to CDR-H3, and the amino acid sequence is shown in SEQ ID NO.6-SEQ ID NO.8; the light chain variable region has three complementary determining regions CDR-L1 to CDR-L3, and the amino acid sequence is shown in SEQ ID NO.9-SEQ ID NO.11. The monoclonal antibody of the present invention can specifically bind to monkey interleukin-6, and the above-mentioned monoclonal antibody is used to screen a pairable monoclonal antibody. Based on the combination of the monoclonal antibodies, a monkey interleukin-6 enzyme-linked immunosorbent assay kit is obtained, which can detect the content of monkey interleukin-6 in monkey serum, plasma or tissue fluid.

This invention discloses an enzyme-linked immunosorbent assay (ELISA) kit for detecting benzo[a]pyrene in edible oil, kitchen fumes, and vehicle exhaust. The kit includes: a sample reconstitution solution, an enzyme conjugate / antibody diluent, a standard / sample diluent, a benzo[a]pyrene standard, a benzo[a]pyrene-specific mouse monoclonal antibody, an enzyme conjugate, an ELISA plate coated with benzo[a]pyrene antigen, a chromogenic solution, a stop solution, and a washing solution. This invention also discloses a method for detecting benzo[a]pyrene in edible oil, kitchen fumes, and vehicle exhaust using the kit, including sample pretreatment, ELISA detection, and result processing and analysis. The sample reconstitution solution, enzyme conjugate / antibody diluent, and standard / sample diluent provided by this invention overcome the solubility problem of benzo[a]pyrene in edible oil, kitchen fumes, and vehicle exhaust while maintaining antibody sensitivity, and can also significantly improve antibody titer, thus possessing significant value for widespread application.

To provide methods for stably producing an antibody enzyme excellent in storage stability.SOLUTION: There is disclosed a method for producing a κ light chain antibody comprising the steps of: expressing a κ light chain antibody in an intracellular or extracellular expression system by using an expression vector comprising a polynucleotide that encodes the κ light chain antibody; subjecting the expressed product to column chromatography using a column containing a first filler to obtain a crude product containing the κ light chain antibody; adding metal ions to the crude product containing the κ light chain antibody so as to obtain a crudely purified product containing a complex of κ light chain antibody with the metal ions; purifying the crudely purified product containing the κ light chain antibody-metal complex by column chromatography using a column containing a second filler from the crudely purified product containing the complex of the κ light chain antibody, to obtain a purified κ light chain antibody-metal complex; and removing the metal ions from the purified κ light chain antibody-metal complex so as to obtain a purified product of the κ light chain antibody.SELECTED DRAWING: None

The present invention relates to the application of styrene-maleic anhydride copolymers and their composites; during the application process, the stability of enzymes and the solid-phase carriers have an impact on antibodies, enzyme activity, and detection. For example, the surface characteristics of magnetic bead particles have an impact on both enzyme activity and detection. Styrene-maleic anhydride copolymers can be used as surface functional groups on chips to bind biological macromolecules and can be used as an ideal photosensitive material in surface plasmon resonance. The present invention can improve the activity and stability of labeled bioenzymes through solid-phase carriers, thereby improving detection sensitivity and reducing detection costs.

The invention relates to the technical field of enzyme-linked immunosorbent assay, in particular to a hapten for creatine content detection, an antigen, an antibody and an enzyme-linked immunosorbent assay kit, and further discloses application of the hapten in the aspects of the antigen, the antibody and the enzyme-linked immunosorbent assay kit. The enzyme linked immunosorbent assay kit established by aiming at the hapten for creatine content detection can realize rapid detection of a large batch of samples to be detected, is convenient to use, low in detection cost, efficient, accurate and rapid in detection method, can simultaneously detect a large batch of samples, and is suitable for creatine content detection.

The application provides a flumetover half antigen, artificial antigen and antibody and a preparation method of the flumetover half antigen, artificial antigen and antibody, and application of the flumetover half antigen, artificial antigen and antibody to an enzyme-linked immunoassay kit, the kit comprising: an enzyme-labeled plate coated with a coating agent, a flumetover standard solution, a flumetover antibody, an enzyme conjugate concentrate, an enzyme conjugate diluent, a substrate developing solution, a termination solution and a washing solution, wherein the coating agent is a flumetover conjugate antigen, the enzyme conjugate is an enzyme-labeled flumetover antibody, and the flumetover antibody is obtained by immunizing animals with an immunogen. The application further discloses a method for detecting flumetover by using the enzyme-linked immunoassay kit, which comprises the following steps: firstly, sample pretreatment, then, detection by using the kit, and finally, analysis of the detection result. The enzyme-linked immunoassay kit provided by the application can be used for detecting the content of flumetover in vegetable and fruit samples, and has the advantages of simple operation, low cost, high sensitivity, on-site monitoring and suitability for screening of a large number of samples.

The present invention belongs to the field of biological detection technology, and specifically relates to an anti-monkey interleukin-6 monoclonal antibody, an enzyme-linked immunosorbent assay kit and its application. Named IL-6-12, IL-6-12 includes a heavy chain variable region and a light chain variable region; the heavy chain variable region has three complementary determining regions CDR-H1 to CDR-H3, and the amino acid sequence is shown in SEQ ID NO.6-SEQ ID NO.8; the light chain variable region has three complementary determining regions CDR-L1 to CDR-L3, and the amino acid sequence is shown in SEQ ID NO.9-SEQ ID NO.11. The monoclonal antibody of the present invention can specifically bind to monkey interleukin-6, and the above-mentioned monoclonal antibody is used to screen a pairable monoclonal antibody. Based on the combination of the monoclonal antibodies, a monkey interleukin-6 enzyme-linked immunosorbent assay kit is obtained, which can detect the content of monkey interleukin-6 in monkey serum, plasma or tissue fluid.

An enzyme-linked immunosorbent assay kit for detecting cannabidiol and its application. The kit includes: an enzyme-labeled plate coated with a coating antigen, a cannabidiol standard solution, a cannabidiol-specific antibody, an enzyme-labeled secondary antibody, a chromogenic solution, a termination solution, a washing solution, and a reconstitution solution. The coating antigen is a cannabidiol-conjugated antigen, and the cannabidiol-conjugated antigen is a conjugate of a cannabidiol hapten and a carrier protein. The cannabidiol hapten prepared in the present invention not only retains the characteristic structure of cannabidiol to the greatest extent, but also has a linker with a suitable structure for conjugation with the carrier protein, allowing the characteristic structure of cannabidiol to be fully exposed to the body as much as possible, enhancing the immune effect, improving the specificity of the antibody, and reducing the cross-reaction with other cannabidiol analogs. The enzyme-linked immunosorbent assay kit of the present invention can be used to detect the content of cannabidiol in industrial hemp samples, is easy to operate, low in cost, high in sensitivity, capable of on-site monitoring, and suitable for screening a large number of samples.

The invention relates to a method for determining kininogenase and a used kit. On one hand, the invention relates to a kit for determining the content of kininogenase in human plasma, the kit comprises a test solution for testing, an antigen standard substance, a capture antibody, an enzyme-labeled antibody and a test method description file which are separately packaged by containers, and the test solution comprises PBS, PBST, a washing solution, a confining solution, a sample diluent, a stop solution, a coating buffer solution, a color developing solution A and a color developing solution B. The invention also relates to a method for determining the content of kininogenase in human plasma by using the kit. According to the kit and the method provided by the invention, the amount of endogenous kininogenase or exogenous kininogenase in a human body can be accurately and effectively measured, and a beneficial tool is provided for diagnosis and treatment of clinical diseases.

Methods for target, biomarker, and patient selection discovery in central nervous system disorders utilizing patient-derived cellular models, spatial proteomics, and machine learning. The method generates neural cells from forebrain regions from induced pluripotent stem cells, performs cell-type specific proteome profiling using antibody-enzyme conjugates and spatial proteome profiling, and applies statistical data augmentation to sparse biological datasets. Machine learning classifiers with SHAP-based feature importance identify ranked biomarkers from mass spectrometry data. The platform enables patient stratification by linking molecular signatures to symptom severity, drug screening through biomarker modulation, and diagnostic applications. Kits comprising antibodies for biomarkers including antibodies for biomarkers identified by the method facilitate implementation. Applications include autism spectrum disorder, rare neurodevelopmental disorders, schizophrenia, epilepsy, Alzheimer's disease, and Parkinson's disease.

The present invention relates to a kit and a detection method for detecting the content of aminopeptidase. The kit includes an aminopeptidase antibody, a blocking solution, a standard product, a labeled aminopeptidase antibody, an enzyme-labeled affinity reaction conjugate, and a chromogenic substrate. Among them, the blocking solution includes 0.5-1 wt.% bovine serum albumin and 3-6 wt.% skim milk powder. Another aspect of the present invention is to provide a detection method for the content of aminopeptidase. The enzyme-linked immunosorbent assay is used to detect the content of aminopeptidase in a sample. Among them, the blocking solution includes 0.5-1 wt.% bovine serum albumin and 3-6 wt.% skim milk powder, and the blocking solution is used as a diluent for the labeled aminopeptidase antibody. The kit and the detection method of the present invention can sensitively, stably and reliably detect the content of aminopeptidase in a substance to be detected, providing a powerful means for controlling the aminopeptidase residue in biological products.

A compound having the structure (I) is provided. The compound is useful as a therapeutic agent in the form of a lipoplex, or in a form of a polyplex, in a form of a lipid nanoparticle, or in a form of a liposome, or in a form of a micelle, or in a form of an emulsion including a drug, vaccine, oligonucleotide, antibody, enzyme, protein, small molecule, or nucleotide.

The invention discloses a method for preparing an anti-Mullerian hormone antibody-enzyme marker, comprising the following steps: maleamidation of alkaline phosphatase, sulfhydrylation of an AMH antibody, crosslinking, blocking, and purification. The preparation method optimizes and improves the conventional step of preparing an enzyme-labeled antibody using a heterobifunctional crosslinker. In the method, ultrafiltration is used to purify the maleamidated alkaline phosphatase, the sulfhydrylated AMH antibody, and the labeled AMH antibody-alkaline phosphatase marker, replacing the conventional purification method of dialysis. This not only eliminates the complicated dialysis process, shortens the preparation time of the enzyme-labeled antibody, but also reduces raw material loss. Also disclosed are the anti-Mullerian hormone antibody-enzyme marker prepared by the preparation method and the use of the anti-Mullerian hormone antibody-enzyme marker in preparing an AMH enzyme-labeled antibody detection reagent. The prepared AMH enzyme-labeled antibody detection reagent has high sensitivity and good stability in detecting AMH.

The present disclosure describes a device and / or composition comprising a hydrogel system, such as an alginate methacrylate hydrogel system, and a therapeutic agent. The device and or composition may contain one or more therapeutic agents, such as an engineered cell or protein, a secreted protein, a hormone, a cytokine, an antibody, an enzyme or a peptide Also described herein is a method of treating a disease or disorder using a device and / or composition comprising a hydrogel system and a therapeutic agent.

The invention discloses a monoclonal antibody combination for Brucella detection and application thereof, and belongs to the technical field of immunodetection. The monoclonal antibody combination disclosed by the invention comprises a monoclonal antibody 15F4 and a monoclonal antibody 32H9. On the basis of the monoclonal antibody combination, a double-antibody sandwich ELISA detection method with 15F4 as a capture antibody and enzyme-labeled 32H9 as a detection antibody is established, and key parameters such as coating concentration, closing conditions, incubation and color development time and the like are optimized. The method has high specificity on brucella, and does not cross with common cross-reactive bacteria such as escherichia coli O: 157, salmonella and the like; the sensitivity is high, and the detection limit can reach 1 * 10 < 5 > CFU / mL; the repeatability is good, and the intra-batch and inter-batch variation coefficients are both smaller than 10%. The invention provides a core reagent and a reliable means for early, rapid and accurate diagnosis of brucellosis, and is suitable for on-site screening of clinical samples and animal epidemic diseases.

The present invention relates to the field of enzyme-linked immunosorbent assay technology, and more particularly to a hapten, antigen, antibody, and enzyme-linked immunosorbent assay kit for detecting creatine content. The present invention also discloses the use of the hapten in the antigen, antibody, and enzyme-linked immunosorbent assay kit. The enzyme-linked immunosorbent assay kit developed for the hapten used to detect creatine content can achieve rapid detection of large quantities of samples to be tested. It is easy to use, has low detection cost, and is efficient, accurate, and rapid. It can simultaneously detect large quantities of samples and is suitable for detecting creatine content.

The invention relates to the technical field of biomedical detection, in particular to a preparation method and application of a biotinylated antibody without purification. The specific quenching agent is added into a reaction system, residual active ester is sealed in situ, so that the detection antibody which can be directly used for ELISA is directly obtained, traditional ultrafiltration or gel filtration purification is omitted, the preparation period is shortened by at least 2-3 hours, and activity loss of the antibody caused by centrifugation or dilution is avoided; the residual NHS-ester is quenched in situ by a quenching agent, so that a background signal is remarkably reduced, and the signal-to-noise ratio is improved (the signal-to-noise ratio in an I-C system is improved by more than or equal to 50%); the method provided by the invention can also be popularized to an antibody-enzyme coupling system of heterobifunctional cross-linking agents such as EMCS, Sulfo-SMCC and the like, and the ELISA performance is equivalent to that of a traditional purified product; the kit is suitable for clinical scenes such as acute myocardial infarction needing rapid diagnosis, and a high-throughput and low-cost detection platform can be conveniently established.