781 results about "Double antibody sandwich" patented technology

The invention discloses a human SARS-CoV-2 monoclonal antibody. The preparation method of the human SARS-CoV-2 monoclonal antibody comprises the steps: adopting SARS-CoV Nucleocapsid recombinant protein as immunogen, immunizing BALB/c mice, performing fusion and subcloning on spleen cells and myeloma cells of mice, then performing a large amount of repeated screening and domestication of cell lines through commercialized products SARS-CoV-2 Nucleocapsid and MERS Nucleocapsid so as to obtain a hybridoma cell line capable of secreting the SARS-CoV-2-resistant N monoclonal antibody with high affinity and high specificity finally and successfully, and finally performing ascites preparation and purification so as to obtain the monoclonal antibody, wherein the amino acid sequence of the SARS-CoVNucleocapsid recombinant protein is shown in SEQ ID No. 1. The invention also discloses application of the monoclonal antibody in preparation of SARS-CoV-2 virus detection products and preparation ofdrugs for inhibiting the SARS-CoV-2 viruses. The monoclonal antibody can be used for detecting the SARS-CoV-2 in human throat swabs/pulmonary secretions and other samples by using a double-antibody sandwich method, and can be applied to diagnosis and prevention and control of SARS-CoV-2 virus infection and scientific researches of viruses and other study.

The invention relates to a kit for detecting neutrophil gelatinase-associated lipocalin content based on chemiluminescence immunoassay. According to the invention, by employing a double-antibody sandwich immunization analysis method, a chemiluminescence magnetic microspheres immunization technology is used, anti-NGAL antibody-coated magnetic microspheres for specifically combining with NGAL antigen of a standard substance/sample in a reaction cup, then are reacted to another strain anti-NGAL antibody labelled with acridine salt to form an immunization compound, through an acid-base chemical reaction of a pre-Trigger and a Trigger, relative light unit (RLU/s) of the chemiluminescence reaction can be measured; the NGAL antigen content in the sample is in direct proportion to the relative light unit (RLU/s) measured by an optical system, determination of NGAL content in an urine specimen can be determined through standard curve fitting; and the method has the obvious advantages of high sensitivity, strong specificity, good stability, simple operation and low cost.

The invention relates to the technical field of biology, and specifically provides a detection kit for a detection kit for antigens of a novel coronavirus (SARS-CoV-2). According to the detection kitprovided by the invention, the antigens of the SARS-CoV-2 are detected by virtue of a colloidal gold double antibody sandwich method, and two monoclonal antibodies and colloidal gold are mixed for labeling, so that a detection result of the antigens of the SARS-CoV-2 is accurate and high in sensitivity, and meanwhile, the detection rate can be remarkably increased. According to the detection kit,a blank in immunological detection of the SARS-CoV-2 is filled up, and field detection can be realized without a detection instrument, so that the time and the labor are saved, and operation is flexible.

The invention provides a homogeneous luminescence immunoassay method for quantitatively analyzing multiple components simultaneously and a kit used for the method. Receptor microspheres containing various different fluoresceins are adopted, and antibody molecules for capturing different biological markers to be measured are enveloped in the method; in a measuring process, the multiple biological markers in a sample to be measured are combined with the corresponding antibody molecules on the surfaces of the receptor microspheres and biotinylated antibodies in a detection system respectively to form double-antibody sandwich compositions which are respectively connected with donor microspheres labeled with streptavidin; when the donor microspheres are irradiated by exciting light, all types of the receptor microspheres send out optical signals with different wavelengths; the intensities of the light with different wavelengths are respectively detected, so that the biological markers to be measured can be accurately quantified. The kit comprises the receptor microspheres containing chemiluminescence reagents and the fluoresceins, the biotinylated antibodies and the donor microspheres containing photosensitive substances. The homogeneous luminescence immunoassay method and the kit have the beneficial effects that simultaneous and quantitative measurement on multiple components is realized, and the detection cost is reduced.

The invention provides a kit for detecting gastrin-17 and a preparation method as well as application thereof. The kit comprises components A and B, wherein the component A is a first anti-gastrin-17 antibody marked with a tracing marker or coated with a magnetic ball, and the component B is a second anti-gastrin-17 antibody coated with the magnetic ball or marked with the tracing marker, or the gastrin-17; moreover, any one of the components A and B is marked with the tracing marker, the other one is coated with the magnetic ball; and the first anti-gastrin-17 antibody and the second anti-gastrin-17 antibody have different binding sites with the gastrin-17. The invention provides a method for detecting the gastrin-17, with the kit provided by the invention, content of the gastrin-17 in a sample can be detected accurately and sensitively according to a double antibody sandwich method principle or a competition method principle.

This invention discloses one EPSPS gene enzyme immune agent case and its use method in the anti-herbicide bean, wherein, the case comprises anti-EPSPS multi-clone antigen, enzyme board with the antigen, enzyme antigens for two, gene switch bean standard product, non-transfer gene bean standard, intense wash liquid, develop liquid and terminal liquid. This invention method comprises the following steps: cloning CP4-EPSPS gene from the gene switch bean expressed in the bacillus coli; then through protein purification complex property to regroup EPSPS protein antibody immune animal to get special single clone antigen and multi-clone antigen; establishing double antigen clamp enzyme immune system test sample to determine its EPSPS protein content.

The invention provides a porcine reproductive and respiratory syndrome virus double-antibody sandwich ELISA kit. The kit comprises: an elisa plate coated with PRRSV N protein monoclonal antibody, an enzyme labeling PRRSV N protein monoclonal antibody, lysis solution and the like. A capture antibody and a detection antibody are respectively aimed at antigenic determinants with different N proteins.The kit provides a reliable means for quick detection of clinical PRRSV antigen. The kit can detects that blood serum only contains 0.2 TCID50 highly pathogenic PRRSV JXwn06 strain (non-highly pathogenic strain can be also be detected). Through detecting clinically collected 80 blood serum samples, compared with RT-PCR result, the specificity of the method is 88 percent, the sensitiveness is 90 percent and the coincidence rate of the specificity and the sensitiveness are 88.8 percent. The kit is convenient for operation, low in use cost, good repetitiveness and suitable for wide promotion andapplication.

The invention discloses a kit and a method for detecting sST2 (soluble ST2) in blood of an abdominal aortic aneurysm and/or aortic dissection patient. According to the kit and the method, a pair of antibodies for identifying different epitopes of sST2 is assembled to obtain a double-antibody sandwich ELISA and immune colloidal gold test strip, so as to realize quantitive and qualitative detection of sST2. Tests prove that the ELISA kit and the immune colloidal gold test strip have relatively high specificity and sensitivity in an abdominal aortic aneurysm marker-sST2 protein of human, are simple to operate, can be utilized for detecting abdominal aortic aneurysm, aortic dissection and other diseases in scientific research and clinical application, and have the functions of auxiliary diagnosis, guide treatment and prognosis judgment.

The invention discloses double-antibody sandwich ELISA (enzyme linked immunosorbent assay) kit used for detecting an avian leukosis group specific antigen. The kit comprises an enzyme plate coated by a monoclonal antibody which is secreted by a hybridoma cell strain the preservation serial number of which is CGMCC (china general microbiological culture collection center) No.5961. The invention also discloses a double-antibody sandwich ELISA method which is established by utilizing the monoclonal antibody and is capable of rapidly and effectively detecting an ALV (avian leukosis virus). In the double-antibody sandwich ELISA method, the monoclonal antibody prepared by pronucleus expressive HLJ09mdj-1p27 albumen is utilized as a peridium antibody, and antibodies prepared by p27 are utilized as a detection antibody. According to the invention, the minimum detection amount of the p27 is 1.25 ng/ml, the method is not reacted with the common virus of birds, and the specificity is good. The method is utilized to detect egg white and an anus swab sample, and the coincidence rate is respectively 96.5% and 88.9% compared with a PCR (polymerase chain reaction) method; and the result proves that the method has the advantages of convenience, celerity, differentia, sensitivity and the like, and is useful for the detection and population purification of the ALV.

The invention relates to an anti-Golgi protein antibody and an application thereof. The invention recombines human GP73 protein immunity animal to obtain an anti-GP73 polyclonal antibody and a monoclonal antibody which specifically aims at GP73 and builds a plurality of methods for detecting GP73 in clinical tissue sections and serum samples, such as immunohistochemical stain and double antibody sandwiched ELISA method and the like. Tests prove that the polyclonal and monoclonal antibodies can be used for preparing a plurality of GP73 detecting agents of different detecting methods.

The invention discloses a CEA TRFIA (time-resolved fluoroimmunoassay) kit based on IMB (immunomagnetic beads). The kit includes a calibrator, immunomagnetic beads connected with CEA antibodies, europium labeled CEA antibodies, an assay buffer solution, a cleaning solution and an enhanced solution. Through double antibody sandwich immunoreaction, IMB-CEA-europium labeled-CEA monoclonal antibody complexes are formed, IMB which adsorb CEA and supernate are separated through magnetic seperation and washed, the enhanced solution is added, and the value is measured through a time-resolved instrument. Besides the TRFIA technology, the invention further has the advantages as follows: through IMB enrichment and full diffusion of IMB in the solution, the combination superficial area is enlarged, the reaction time is greatly shortened, and the detection sensitivity is improved. IMB and antibodies are directionally connected through chemical groups, so that the consumption of paired antibodies is greatly reduced and the detection precision is improved. The technology realizes automation easily, and overcomes the problem that the traditional micropore plate type TRFIA technology can only carry out detection after samples are accumulated to a certain number, thereby realizing real-time sample detection.

The invention belongs to the cancer detection technology, relating to a lung cancer marker detection immunochromatographic test paper and an application. The detection of a single tumour marker in the prior art can not meet the requirements of clinical early diagnosis and differential diagnosis. The invention realizes the early diagnosis of lung cancer by jointly detecting NSE and CEA in serum with a double antibody sandwich immunochromatography. The invention comprises the following preparation steps: preparing aurosol-neure NSE and aurosol-carcinoembryonic antigen CEA antibody compounds; preparing double antibody sandwich NSE and CEA detection test strips. The invention comprises the following steps in lung cancer detection: configurating series concentration NSE and CEA standard substances; preparing NSE and CEA mixed antigen solution with serum of normal people; inserting the sample pad ends of the test strips into the above concentration series standard substances respectively to observe the changes in colours of T and C lines on a NC film. The invention has the advantages of convenient detection operation, high sensitivity and precise result, and can determine lung cancer recurrence 4-12 weeks ahead.

The present invention relates to a dry-type immunoassay test strip and a preparation method and application thereof. On a plastic substrate of the test strip is sequentially stuck with a sample pad, a nitrocellulose membrane anda water absorption pad from left to right, wherein the right end of nitrocellulose membrane is provided with a control line and a detection line that are parallel to each other; the left end of nitrocellulose membrane is provided with a tracer particle labeled antibody/ antigen-coated line and a closed protein line I, wherein the closed protein line I is formed by coating closed protein solution I with a nitrocellulose membrane, and the antibody/ antigen-coated line is formed by spraying the tracer particle labeled antibody/ antigen on a closed protein line II, and the closed protein line II is formed by coating closed protein solution II with a nitrocellulose membrane. . The test strip can be applied in the immunoassay based on the double antibody sandwich method or the competitive method, and has advantages of simple operation, rapid chromatography, high specificity, accurate test results and strong stability.

The invention adopts double-function gold magnetic nanoparticles and provides a detection method which integrates an immunomagnetic bead capturing technology and an immunochromatography technology and is used for rapidly detecting salmonellas. Immunomagnetic separation and immunochromatography are organically integrated, the step of eluting the salmonellas from immunomagnetic beads is eliminated and the capture efficiency is improved; the step of spraying colloidal gold on a bonding mat is eliminated, the immunological reaction is more uniform and a variable coefficient is small in the quantitative detection; workload and probability of mixed bacterium pollution are reduced. The detection method adopts the basic thinking of exploring the mode of effectively combining the gold magnetic nanoparticles and an antibody, optimizing the conditions of enriching the salmonellas and carrying out rapid quantitative detection by using the immunochromatography technology as a vector and using the double antibodies sandwich as a detection principle.

The invention discloses a quick African swine fever virus detection card and application thereof, so as to solve the technical problem of hard African swine fever virus detection. Polyclonal antibodies PoAbI against African swine fever virus p30, P54 and p72 protein labeled with colloidal gold are absorbed on a colloidal gold labeling pad of the detection card; a detection membrane is provided with goat or rabbit anti-mouse IgG antibodies or a quality control line C printed by staphylococcus aureus SPA; and a detection line T printed by polyclonal antibodies PoAbII against African swine fevervirus p30, P54 and p72 protein is also arranged. The invention also discloses a using method for the quick African swine fever virus detection card. The method comprises steps: a to-be-detected sampleis acquired, and PBS or water is added for suspension or grinding; the sample is dripped to the sample loading end of the quick African swine fever virus detection card; and horizontal placing is carried out to observe a result. The quick African swine fever virus detection card adopts a double antibody sandwich method, and has the advantages of strong specificity, high sensitivity, high detection efficiency, high efficiency and practicability, and important practical application value.

The invention discloses a kit for rapid detection of staphylococcus aureus in a sample and a detection method thereof, belonging to the technical field of immunological detection. In the invention, a staphylococcus aureus immunogen inactivated with formaldehyde is used for immunizing a healthy New Zealand rabbit to obtain a polyclonal antibody to serve as a coated antibody, and used for immunizing a BALB/C mouse and performing cell fusion to obtain a monoclonal antibody to serve as a secondary antibody, thus, the kit for performing a double antibody sandwich enzyme-linked immunosorbent assay on the staphylococcus aureus in food (milk) is established, and a rapid and efficient detection means is provided for residual detection of the staphylococcus aureus in the food, and the advantages of lower cost and better stability and repeatability are achieved. A detection limit of the kit is 105cfu/mL and is suitable for detecting mass samples.

The invention discloses a cardiac troponin kit based on a microfluidic chip, a preparation method thereof and a detection method of cardiac troponin. The preparation method comprises the steps of (1)coating the microfluidic chip; (2) performing fluorescent microsphere labeling; (3) assembling the microfluidic chip; (4) preparing a calibration article; (5) acquiring the cardiac troponin cTn1 kit based on the microfluidic chip. The cardiac troponin cTn1 kit performs measuring by means of a double-antibody sandwich method; high-sensitivity time-resolved phosphor is selected as a marker to perform antibody marking on fluorescent microspheres; analytical detection is performed via antibody pair immunoreaction; the performance of a reagent prepared herein may reach the level of same chemical light-emitting reagents. After immunoreaction, a washing liquid is used to fully remove surplus components; detecting and reading are performed on the fluorescent microspheres under immunoreaction bonding; the problem is avoided that the introduction of a developing liquid into the chip causes insufficiency of a chip or failure of timely post-rendering reading.

The invention provides a human N-terminal pro-B-type natriuretic peptide (NT-proBNP) immunoassay kit, mainly comprising 1) a human NT-proBNP calibration material; 2) a vector coated with human NT-proBNP polyclonal antibodies; 3) an enzyme-labeling human NT-proBNP monoclonal antibody; and 4) an enzymatic chemical luminous substrate. The invention further provides a preparation method of the kit. On the basis of the kit of the invention, the enzyme-labeling human NT-proBNP monoclonal antibody, the human NT-proBNP polyclonal antibodies coated on the vector and human NT-proBNP antigens form a 'double-antibody sandwich' structure, thus effectively utilizing a chemiluminescence technical principle and guaranteeing detection sensitivity. In addition, the kit can be applied to an open chemiluminescence measuring instrument; and the kit has low use cost and easier popularization and application.

The invention discloses a preparation method of a multifunctional test paper for early pregnancy. The preparation method comprises the steps: preparing a nitrocellulose membrane; labeling and curing a polyester fiber strip; and assembling and cutting. A human chorionic gonadotropin beta core segment and a human chorionic gonadotropin regular molecule in urine are detected by applying a principle of a double-antibody sandwich method and an immunochromatographic method, detection steps are simple, a result can be observed within 5 min, and a common user can detect the result, and the detection result is more accurate. Through comparing color depth of a first detection line and a second detection line, whether to be pregnant and whether to be normally pregnant are determined, and ectopic gestation and threatened abortion are subjected to risk assessment.

The invention discloses combined detection test paper of influenza A virus antigen and influenza B virus antigen and a preparation method thereof. The test paper comprises a sample pad, a fiberglass membrane containing a colloidal gold particle label, a nitrocellulose membrane and water absorbing paper, wherein the nitrocellulose membrane comprises a detection area which is coated with an influenza A virus antibody, a detection area which is coated with an influenza B virus antibody and a control area which is coated with an goat anti-rabbit antibody; the colloidal gold particle label comprises a micro signal amplification system and a colloidal gold labeled rabbit IgG antibody; and the micro signal amplification system is a colloidal gold particle-avidin-biotin-influenza A/B virus antibody. According to the invention, an avidin-biotin microsignal amplification system is added in a double-antibody sandwich detection system, the signal of a target antibody is enlarged, the detection sensitivity is increased, false negative or detection omission due to weak signals can be avoided, simultaneously combined detection can be carried out on the influenza A and B virus antigens, and the detection time, sample and cost can be saved.

The invention discloses an on-site detection immuno-chip, comprising a chip carrier and an agarose gel layer which is paved on the chip carrier, wherein, the agarose gel layer is fixed with a plurality of antibody microarrays of 4*4, and chip-dedicated fences or Super PAP Pen scribings are used for separating the microarrays from each other; the double antibody sandwich principle is adopted to detect the antigen to carry out the dot matrix of corresponding capture antibody, positive control and negative control on the solid phase carrier simultaneously; the antibody protein is connected with the solid phase carrier through the covalent bond and physical adsorption; the sample liquid to be detected and the chip are directly incubated; the antigen to be detected in the samples combine with the corresponding antibody which is fixed on the chip; a specific monoclonal antibody probe marked by horse radish peroxidase is added and the macroscopic detection results can be obtained after the coloration of substrates. The invention can detect viruses in multiple aquatic animal samples and the results are macroscopic, thus being applicable to rapid and accurate detection of viruses of aquatic animals in breeding production.