187 results about "Immunoassay method" patented technology

Provided is an immunological measurement method for measuring a target substance in a sample, wherein an antigen-antibody reaction is performed at least once in the presence of an enzyme that specifically decomposes immunoglobulin A, abbreviated as IgA.

The present invention is directed to assay methods and compounds and kits used in such assays.

The invention discloses a YOLO-Drop-based 3D printing droplet microfluidic immunoassay method, which organically integrates a high-precision microfluidic chip preparation technology, a droplet-level immunoreaction system and a fluorescence imaging acquisition and deep learning detection algorithm. And full-chain closed-loop processing from droplet generation, fluorescence signal acquisition, image recognition, result statistics to concentration quantitative analysis is realized. Through deep combination of chip physical parameters and model input features, the method not only significantly improves the accuracy and stability of droplet identification and concentration quantification, but also enhances the robustness of the algorithm under complex sample conditions through multi-scale feature fusion and a signal dual screening mechanism. Meanwhile, the optimized YOLO-Drop model supports embedded efficient reasoning, so that the whole analysis process has the characteristics of real-time performance and high throughput, and the method is suitable for rapid quantitative detection of various biomarkers.

The present invention relates to a multiplex immunoassay method for diagnosing autoimmune nodopathy and, more specifically, to a multiplex immunoassay method, comprising the steps of: (1) binding CNTN1, Caspr1, NF155, and NF186 proteins each to microspheres having different color codes; (2) mixing the microspheres having different color codes to which the proteins are each bound; (3) treating the mixed microspheres with a biological sample to induce an antigen-antibody reaction; (4) adding a detection antibody; and (5) reading fluorescence information of the microspheres to measure the presence or absence of anti-CNTN1 antibody, anti-Caspr1 antibody, anti-NF155 antibody, or anti-NF186 antibody in the biological sample and quantify amounts thereof, thereby enabling simultaneous testing of four antibodies for rapid diagnosis of autoimmune nodopathy.

The invention provides a safrole hapten and application of the safrole hapten in detection of safrole through immunoassay. The structure of the safrole hapten is shown in the specification. The two safrole haptens provided by the invention have spacer arms with proper lengths, and after the safrole haptens are coupled with carrier protein, the haptens can be fully exposed, and recognition of the spacer arms can be avoided as much as possible. The safrole hapten is used for preparing an artificial antigen and an antibody, and the obtained safrole monoclonal antibody is high in titer, strong in specificity and high in affinity. An immunoassay method constructed on the basis of the safrole artificial antigen and antibody has high specificity and sensitivity, the lowest detection limit LOD is 0.09 ng / mL, the half inhibitory concentration IC50 is 2.05 ng / mL, the quantitative detection range is 0.28-14.89 ng / mL, no cross reaction exists on safrole analogues, and rapid qualitative and quantitative detection can be conducted on safrole in a sample.

The preparation and evaluation method comprises the following steps: 1, extracting a heavy-chain DNA fragment and a light-chain DNA fragment, carrying out recombinant transformation by using an overlapping PCR method to construct an scFv gene, 2, constructing an expression vector, culturing the expression vector, carrying out sequencing identification to obtain an scFv bacterial solution, recombining the scFv bacterial solution with a mouse Fc fragment, and identifying a sequence to obtain an MN8 bacterial solution, the method comprises the following steps: 1, preparing an MN8 bacterial liquid, 2, extracting the MN8 bacterial liquid, 3, carrying out inoculated culture on the MN8 bacterial liquid to obtain an MN8 genetically engineered antibody bacterial liquid, 4, extracting MN8 genetically engineered antibody bacterial liquid plasmids and carrying out transfection expression purification, and 5, carrying out antibody titer and thermal stability determination on the MN8 genetically engineered antibody. The scFv single-chain antibody obtained by recombination according to the method has the advantages of small molecular weight, high penetrability and the like, the stability and the sensitivity of an immunoassay method of the scolosporins can be improved, and a certain basis is provided for rapid determination of the scolosporins.

The invention belongs to the technical field of immunodetection, and discloses an avian single-chain antibody for detecting a liver cancer marker GPC3, a kit and a detection method. The poultry-derived single-chain antibody has the characteristics of small volume, high specificity, high stability and good sensitivity, and can be used in an immunoassay method for detecting a liver cancer marker GPC3; according to the method, a chicken-derived single-chain antibody is coated on a solid-phase carrier, a rabbit-derived polyclonal antibody is taken as a detection carrier, the antibodies are combined with a GPC3 antigen to form a sandwich structure, and the double-antibody sandwich immunoassay method of the GPC3 is established by using the method, and has the advantages of high specificity, good sensitivity, high sensitivity and high sensitivity. The invention provides a rapid, accurate and reliable detection method for detecting GPC3 in serum.

The invention provides an analgin hapten, a preparation method thereof, an artificial antigen, an antibody and application. The structures of the analgin haptens are as shown in a formula (I) or a formula (II), the two analgin haptens provided by the invention have spacer arms with proper lengths, and after the two analgin haptens are coupled with protein, the haptens can be fully exposed to an immune system, the spacer arms can be prevented from being recognized as much as possible, and a better antibody is generated to recognize analgin. The analgin hapten is used for preparing artificial antigen and antibody, the obtained analgin monoclonal antibody has high titer and strong specificity, an immunoassay method constructed based on the analgin monoclonal antibody has high specificity and sensitivity, analgin in a sample can be rapidly, qualitatively and quantitatively detected, the operation is simple and convenient, and the detection result is accurate and reliable.

The invention discloses an ultrasensitive immunodetection method based on adjacent nucleic acid amplification signal amplification in the technical field of medical detection. The ultrasensitive immunodetection method comprises the following steps: step 1, immune binding; step 2, enzyme digestion release: releasing a hairpin loop DNA single strand; step 3, complementation and enzyme ligation: adding T4 ligase to cyclize the circular DNA single chain; step 4, rolling circle amplification; and step 6, detecting, and quantifying and positioning by qPCR (quantitative polymerase chain reaction) and an in-situ oligonucleotide fluorescence labeling detection method. According to the present invention, the hairpin ring structure on the oligonucleotide DNA chain coupled to the antibody protects the complementary sequence so as to avoid the generation of the non-specific circular DNA single chain, and the rolling circle amplification technology is combined to amplify the detection signal, such that the detection background signal intensity is inhibited while the specificity of the detection signal is enhanced so as to further improve the detection sensitivity.

The invention belongs to the technical field of immunodetection, and discloses a preparation and characterization method of a rabbit-derived single-chain antibody for detecting a liver cancer marker GPC3. The rabbit-derived single-chain antibody has the characteristics of small volume, high specificity, high stability and good sensitivity, and can be used in an immunoassay method for detecting a liver cancer marker GPC3; according to the method, a specific rabbit single-chain antibody is obtained through screening, expression and purification by using a phage display technology, a GPC3 antigen is coated on a solid-phase carrier, the rabbit single-chain antibody is used as a detection carrier, the binding force of the single-chain antibody and the antigen is identified, and the method is high in specificity, good in sensitivity and high in specificity. A novel single-chain antibody with high specificity and high sensitivity is provided for detection of GPC3 in serum.

Described herein are immunoassay methods for detecting and / or monitoring a cancer in a patient. In the method a biofluid sample from a patient is contacted with a monoclonal antibody that specifically binds to a C-terminal epitope of type XXVIII collagen, and the amount of binding between the monoclonal antibody and peptides in the sample is detected and determined.

The invention comprises a micro-fluidic chip and an immunodetection method. The micro-fluidic chip comprises a reaction layer substrate and a micro-channel sample adding layer positioned on the reaction layer substrate; the reaction layer substrate comprises a base, a three-electrode part prepared on the base and a contact metal pad connected with an external circuit, and the three-electrode part comprises a working electrode, a reference electrode and a counter electrode; a plurality of micro-channel grooves are formed in the bottom of the micro-channel sample adding layer, a plurality of up-down through inflow holes and outflow holes are formed in the micro-channel sample adding channel layer, and the two ends of each micro-channel groove are communicated with the inflow holes and the outflow holes respectively. The micro-fluidic chip is designed through photoetching, PVD and other technologies, the chip comprises a plurality of micro-channels, interference of the external environment to the detection process can be reduced through testing in a closed environment, and the micro-fluidic chip has the rapid, stable and high-throughput detection function and can be applied to bedside detection.

Described herein are immunoassay methods ("PRO-C17 assay") for measuring the extracellular domain levels of XVII collagen present in a patient sample, which can be used to detect and / or monitor cancer and / or assess the severity of cancer in a patient. Monoclonal antibodies and assay kits suitable for carrying out the methods are also described.

The present invention discloses an anti-sulfadiazine nano-antibody and its application, belonging to the field of nano-antibody technology. In order to provide a nano-antibody for detecting sulfadiazine that has a small molecular weight, strong penetrating power, good stability, strong antigen binding ability, and can recognize antigen epitopes that are difficult for conventional antibodies to recognize. The present invention provides a nano-antibody for detecting sulfadiazine, the amino acid sequence of CDR1 of the variable region of the nano-antibody is shown as SEQ ID NO.6, the amino acid sequence of CDR2 of the variable region of the nano-antibody is shown as SEQ ID NO.7, and the amino acid sequence of CDR3 of the variable region of the nano-antibody is shown as SEQ ID NO.8. The indirect competitive ELISA method established by the present invention is a simple, rapid, sensitive, and low-cost immunoassay method, which has great application prospects in the detection of sulfadiazine residues in animal-derived foods.

Disclosed are: a monoclonal antibody which enables the rapid, simple, and highly sensitive detection and measurement of SARS-COV-2 contained in a test sample; and an SARS-COV-2 immunoassay method and immunoassay instrument using the monoclonal antibody. A monoclonal antibody or an antigen-binding fragment thereof comprises a heavy chain CDR1-CDR3 having a specific amino acid sequence, and a light chain CDR1-CDR3 having a specific amino acid sequence. The immunoassay method for the SARS-COV-2 comprises the step of performing immunoassay on the SARS-COV-2 by utilizing the monoclonal antibody or the antigen binding fragment thereof to react with the antigen-antibody of the SARS-COV-2 in a test sample.

This disclosure provides an immunoassay with exceptional sensitivity for detecting a target antigen in a sample. In one disclosure, the capture and detection steps of such an assay utilize recombinant antibodies. The disclosed immunoassay may find specific applications in the detection of HBV antigen.

Disclosed are an amyloid β measurement method and a reagent capable of more accurately measuring amyloid β and unaffected by molecular target drugs (antibody drugs) for amyloid β. Provided is an immunoassay method for amyloid β in a sample isolated from a living body including a pretreatment step for mixing a sample isolated from a living body, (1) an acidifying agent, and (2) at least one selected from the group consisting of cationic surfactants, zwitterionic surfactants, protein denaturants, and chelating agents.

The present invention belongs to the field of immunochromatographic detection technology, and specifically relates to a colorimetric immunochromatographic detection probe, a test strip, a detection method and an application. The detection probe is a magnetic mesoporous metal nanomaterial probe, which includes a magnetic material, a mesoporous silicon material, a dopamine hydrochloride flexible support material and precious metal nanoparticles. The mesoporous silicon material is located between the core of the magnetic material and the dopamine hydrochloride flexible support material, and the precious metal nanoparticles are adsorbed on the surface and pores of the mesoporous silicon material. The surface of the dopamine hydrochloride flexible support and the precious metal nanoparticles is enriched with detection antibodies. The precious metal nanoparticles coated on the colorimetric immunochromatographic detection probe of the present invention can quickly trigger the gold plasma surface resonance effect, so that Au 3+ The reduced gold particles are deposited on the chromatographic test strip, thereby enhancing the colorimetric intensity. Notably, adjusting the pH of the hydroxylamine solution effectively reduces false-positive interference caused by self-nucleation. Therefore, this immunoassay method enables rapid, ultrasensitive, and visual detection of biomarkers.

To provide a small, easy-to-operate electrochemiluminescence sensor. [Solution] An electrochemiluminescence sensor for detecting a test substance in a sample using nanoparticles with an electrochemiluminescence immunoassay method utilizing a plasmon-enhanced field, comprising: a first substrate; an electrode provided on the surface of the first substrate, wherein a first specific binding substance that specifically binds to the test substance is bound to the surface of the electrode; the nanoparticles comprising: metal nanoparticles; a polymer film covering the surface of the metal nanoparticles; a second specific binding substance bound to at least one of the polymer film and the metal nanoparticles and specifically binding to the test substance; and an electrochemiluminescent substance bound to at least one of the polymer film and the second specific binding substance and capable of contacting a co-reactant, wherein the electrochemiluminescence sensor comprises: a first substrate; an electrode provided on the surface of the first substrate, wherein a first specific binding substance that specifically binds to the test substance is bound to the surface of the electrode; and an electrochemiluminescent substance that is bound to at least one of the polymer film and the second specific binding substance and capable of contacting a co-reactant.

The present invention relates to an immunoassay device and an immunoassay method.According to an aspect of the present invention, an immunoassay device includes a measurement unit provided with a detection unit disposed at one side of a stage accommodating cartridges having a plurality of wells to move in a direction, in which the plurality of cartridges are arranged, and capable of measuring a state within the well disposed at the outermost side, and including a shielding plate that moves to cover an opened upper portion of the well disposed at the outermost side to block introduction of light into the well.

The presently described and claimed technology relates immunoassay methods of assessing a subject's Alzheimer's Disease status and zygosity for the APOE ε4 allele.

This invention belongs to the field of immunology and discloses a hybridoma cell line and the preparation and application of its secreted monoclonal antibody that uniformly recognizes tylosin / tilmicosin. The monoclonal antibody hybridoma cell line is named DES-AOAA-14D5, with accession number CGMCC NO: 45303. This invention utilizes decarboxymethyl tylosin and oxycarboxymethyl hydroxylamine to synthesize a hapten, conjugates a carrier protein to prepare a complete antigen, immunizes mice, and, through cell fusion and screening techniques, prepares a hybridoma cell line that can secrete a monoclonal antibody that uniformly recognizes tylosin / tilmicosin. IC50 50 The effective concentrations were 1.59 and 1.72 ng / mL, respectively, with a cross-reactivity rate as high as 92.44%. An immunoassay method for the simultaneous and accurate detection of tylosin / tilmicosin in milk was established using this antibody. The cut-off value for both tylosin and tilmicosin was 16 ng / mL, and no cross-reactivity was observed with other structural analogs. The detection method of this invention shows promising application prospects.

The invention provides a feline herpesvirus antibody fluorescent microsphere detection test paper and a preparation method thereof, FHV gD protein is used as a specific capture antigen coating T line, feline IgG is used as a C line coating object, SPA and Eu (III) chelate particles are combined based on a lateral chromatography immunoassay method, and the fluorescent microsphere detection test paper for the feline herpesvirus antibody is obtained. The T-line fluorescence value, the C-line fluorescence value and the T / C value of the fluorescent microsphere detection test paper for the feline herpesvirus antibody are improved, accurate, rapid and quantitative indirect detection of the FHV antibody in feline serum is realized, and the sensitivity and the specificity of the detection test paper are improved. By testing a cat serum sample and a clinical sample, the detection result is consistent with that of a traditional standard method, and the reliability and effectiveness of the method in the aspect of on-site monitoring of the FHV antibody in the cat body are proved. Compared with a colloidal gold immunoassay method, generation of false positive results is avoided, and a feasible solution is provided for timely diagnosis and control of feline herpesvirus infection.

Provided is a heterohybridoma-based method of generating recombinant rabbit monoclonal antibodies, recombinant anti-IL-6 receptor-alpha (IL-6Rα) antibodies generated using such methods, and immune assay methods kits employing such antibodies.

Disclosed herein are methods of immunoassay for detecting in a patient sample (such as for example a blood, serum or plasma sample) a biomarker (PRO-C12) originating from Type XII Collagen. Said methods may be used for detecting and / or monitoring cancer or a level of severity thereof. Also disclosed are monoclonal antibodies and assay kits for use in said methods of immunoassay.

The invention provides a linear epitope of an African swine fever virus CP312R protein. The amino acid sequence of the linear epitope is SEQ ID NO: 1 or SEQ ID NO: 2. The antigen epitope polypeptide provided by the invention has good antigen specificity and immunoreactivity. The polypeptide can be efficiently combined with an African swine fever virus specific antibody, background interference is low, cross reaction is small, and detection sensitivity and accuracy are remarkably improved. By using the polypeptide as a coating antigen in immunodetection methods such as ELISA or colloidal gold immunochromatography, rapid and accurate detection of ASFV infection can be realized, and the detection result is stable and reliable. Compared with a traditional whole virus antigen or a crude extract antigen, the epitope polypeptide has the advantages of being simple and convenient to prepare, small in batch difference, high in safety and the like, and is suitable for ASFV serological monitoring and diagnosis application.

The application discloses an antibody for detecting phosphorylated Tau protein pTau217, an immunodetection method and application. The antibody comprises a T217-3D6 antibody; light chain CDR1, CDR2 and CDR3 of the T217-3D6 antibody are sequentially the sequences shown in SEQ ID NO. 1 to 3, and heavy chain CDR1, CDR2 and CDR3 are sequentially the sequences shown in SEQ ID NO. 4 to 6. The antibody is used for immunodetection of the phosphorylated Tau protein pTau217, and has the advantages of simple operation, high sensitivity and strong specificity, and can realize rapid detection of the phosphorylated Tau protein pTau217, and has important significance for evaluating the Tau protein phosphorylation level and Tau protein pTau217 phosphorylation related detection.

This invention belongs to the field of molecular biology detection technology and is mainly used to solve the problem of large amounts of washing solution consumption caused by frequent rinsing in solid-phase immunoassay methods. Specifically, based on the characteristic that the artificial introduction of upstream reagents in solid-phase immunoassay methods will not affect downstream steps, the liquid in the upstream rinsing step is recovered and used for rinsing downstream steps. To avoid cross-binding of upstream analyte-specific reagents between different samples, the recovered washing solution is used for downstream rinsing using an instantaneous rinsing method to minimize the time spent in the process. When the entire batch of samples is testing for the same analyte, the recovered washing solution can be used for immersion washing in each rinsing step.

The present invention relates to immunoassay methods suitable for detecting and / or monitoring cancer in patients, immunoassay kits suitable for carrying out said methods, and antibodies suitable for use in said methods and kits.

This invention provides an immunoassay method comprising the following steps: S1: performing two parallel immunoassays on a sample containing the target molecule, recording the results of the two parallel immunoassays as the first and second values, respectively; S2: calculating the ratio of the first value to the second value; S3: determining the concentration of the target molecule in the sample. The assay method of this invention can solve the problem of HOOK effect samples, and the method is not limited by the detection range, capable of directly measuring up to 10... 6 High sample concentrations at the ng / ml level; good repeatability and fast detection speed.