31 results about "Immune adsorption" patented technology

The invention provides a method for detecting a virus inactivation effect by an ELISA (Enzyme-Linked Immunosorbent Assay) method, which comprises the following steps: acquiring a to-be-detected inactivated virus sample, extracting an antigen component from the sample, and detecting an antigen structure change characteristic value through a high-throughput mass spectrometry analysis technology to obtain antigen degradation degree data; obtaining a sample subjected to preliminary inactivation, and detecting the antigen content change trend through an enzyme-linked immunosorbent assay to obtain content change curve data; detecting the residual quantity of viral nucleic acid by a real-time fluorescent quantitative PCR (Polymerase Chain Reaction) technology aiming at the immunogenicity retained sample to obtain nucleic acid degradation degree data; acquiring nucleic acid degradation degree data, classifying the relationship between nucleic acid residues and infectivity by adopting a support vector machine algorithm, and judging that the inactivation effect is completely completed if the classification result shows that the infectivity is lost; according to the antigen content change trend, the immunogenicity index and the nucleic acid degradation degree data, a weighted fusion algorithm is adopted to integrate multi-dimensional detection results, and a comprehensive inactivation effect score is obtained.

The present disclosure relates to the use of DNA-peptide hybrid molecules to detect target molecules in a sample. In some embodiments, the DNA-peptide hybrid molecules comprise target-specific binding peptides which selectively bind to a target molecule. Kits comprising DNA-peptide hybrid molecules are also provided.

The invention relates to a detection method, in particular to a method for rapidly detecting alpha and beta-lactoglobulin in auxiliary material lactose based on a microarray chip technology. On the basis of an indirect competitive enzyme-linked immunosorbent assay principle and a silver enhanced developing technology, a high-sensitivity microarray chip detection platform is successfully constructed by virtue of a biological chip scanner, and rapid quantitative analysis of alpha-lactalbumin and beta-lactoglobulin in the auxiliary material lactose for injection is realized. According to the method, an antigen-antibody specific recognition mechanism and a nano-silver signal amplification mechanism are integrated, and the detection efficiency and sensitivity are remarkably improved through the high-throughput layout of the microarray chip. In the aspect of methodology performance, the technology has the remarkable advantages of simple and convenient operation process, short detection time, high sample treatment flux, excellent specificity (CRlt; 1%) and the like.

The invention discloses an anti-Ebola virus VP40 protein nano antibody and application of the anti-Ebola virus VP40 protein nano antibody in Ebola virus detection. The anti-Ebola virus VP40 protein nano antibody is a 3E07 antibody. The anti-Ebola virus VP40 protein nano antibody prepared by the invention has the characteristics of high sensitivity, wide binding spectrum and the like. A double-antibody sandwich enzyme-linked immunosorbent assay method constructed by the 3E07 and 3F06 antibodies can be used for efficiently detecting VP40 proteins and virus-like particles of various subtype Ebola viruses such as Zaire, Sudan, Bandong and the like, and the detection sensitivity for the VP40 proteins of the Zaire type Ebola viruses can reach 0.039 ng / mL; the antibody combination has a wide application prospect in diagnosis and pathogen detection of Ebola virus diseases.

The invention belongs to the technical field of biological medicines, and particularly relates to a dissociation liquid applied to an aluminum adjuvant adsorption influenza vaccine and a method for measuring the antigen content. The dissociation liquid disclosed by the invention is prepared from a potassium phosphate salt solution, diethanol amine, TritonX-100, tween-20, dodecyl dimethyl betaine and water. The dissociation liquid is excellent in dissociation efficiency, the dissociation recovery rate of the dissociation liquid to H1N1, H3N2 and BV type influenza vaccine hemagglutinin reaches 106%-110%, the antigen activity can be effectively protected, operation is easy and convenient, adaptability is wide, a dissociated sample can be directly used for mainstream detection methods such as a one-way immunodiffusion test, an enzyme-linked immunosorbent assay and a high performance liquid chromatography method, and the dissociation liquid has good application prospects. The precise quantification of the antigen content in the aluminum adjuvant adsorption influenza vaccine is realized, and a reliable technical support is provided for vaccine quality control.

The invention relates to key antigen epitope screening, in particular to an echovirus 18 type key antigen epitope peptide and application thereof. The result of an enzyme-linked immunosorbent assay shows that Epitope 1 (SEQ ID NO: 1) can be effectively combined with E18-VP1 antiserum, and the combining capacity is obviously higher than that of Epitope 2 in a blank control and a negative result, so that one antigen epitope peptide is screened based on the complete sequence of the echovirus 18-VP1, and the antigen epitope peptide has important guiding significance for research and development of echovirus 18 vaccines.

The application provides a screening method of human amniotic membrane mesenchymal stem cells (hAMSCs) with bone regeneration promoting potential, and uses the protein level of transmembrane protein 119 (TMEM119) as a functional index of the bone regeneration promoting ability of the hAMSCs, and uses an enzyme-linked immunosorbent assay (ELISA) to detect the TMEM119 protein concentration in a lysis solution prepared from 1×10 7 The application uses the TMEM119 protein concentration not lower than 15000 pg / mL as a screening standard of the hAMSCs with bone regeneration promoting potential, and screens the hAMSCs with significant bone regeneration promoting potential from different batches of hAMSCs, so that the hAMSCs meeting the screening standard of the application have obvious bone formation promoting potential, and the technical problem that the hAMSCs with bone regeneration promoting potential are difficult to be accurately identified and screened in clinical transformation is solved.

The invention discloses an application of nitric oxide synthase neurons in an island leaf cortex II layer in preparation of a medicine for treating osteoporosis, and systematically reveals a nerve stress induced bone metabolism imbalance mechanism by constructing a chronic heterologous pressure stress mouse model and combining an enzyme-linked immunosorbent assay and a high-resolution micro-CT (Computed Tomography) technology; the method comprises the following steps: accurately positioning specific nerve connection between nNOS neurons of an island leaf cortex II layer and bone tissues for the first time by adopting a reverse cross-polysynaptic virus tracing technology, and determining that the region is a key central target point regulated and controlled by a brain-bone axis; the activity of target neurons is specifically intervened through a chemical genetics technology, it is proved that activation can induce the phenotype of osteoporosis, and pathologic indexes are remarkably improved through inhibition.

The present invention discloses a camel-derived single-domain antibody GA4 that recognizes a linear B-cell antigen epitope in Clostridium difficile glutamate dehydrogenase and its application. The nucleotide sequence of the camel-derived single-domain antibody GA4 is shown in SEQ ID NO: 1, and the amino acid sequence is shown in SEQ ID NO: 2. The camel-derived single-domain antibody GA4 of the present invention has the advantages of strong specificity, high sensitivity, and good accuracy. It has been demonstrated that it can be used for detection of recombinantly expressed CD-GDH and endogenous CD-GDH by methods such as enzyme-linked immunosorbent assay (ELISA) and western blot hybridization (WB).

The invention discloses a camel source single-domain antibody CB3 for recognizing linear B cell epitope in clostridium difficile methylase CamA and application of the camel source single-domain antibody CB3. The nucleotide sequence of the camel source single-domain antibody CB3 is as shown in SEQ ID NO: 1, and the amino acid sequence of the camel source single-domain antibody CB3 is as shown in SEQ ID NO: 2. The camel source single-domain antibody CB3 disclosed by the invention has the advantages of strong specificity, high sensitivity, good accuracy and the like, and it is proved that the camel source single-domain antibody CB3 can be used for detecting recombinant expressed CD-CamA and endogenous CD-CamA by methods such as enzyme-linked immunosorbent assay (ELISA) and western blot hybridization (WB).

The present invention discloses a hapten for detecting toxoflavin, and an artificial antigen and antibody for detecting toxoflavin are prepared based on the hapten. The hapten completely retains the structural characteristics of toxoflavin, and the artificial antigen prepared by the hapten has an intact structure and completely retains the original characteristics of toxoflavin, which is conducive to presenting the structural characteristics of toxoflavin and improving the immunogenicity of the artificial antigen. The artificial antigen can quantitatively detect toxoflavin in a sample to be tested. When used in enzyme-linked immunosorbent assay (ELISA) of toxoflavin, IC 50 The value is 0.41μg / L; at the same time, based on the artificial antigen, an immunochromatographic test strip for the quantitative detection of toxoplasmone is provided. The detection sensitivity of toxoplasmone standard reaches 0.1μg / L, and the detection limit of toxoplasmone in the test sample reaches 25ng / kg, which can achieve high-sensitivity detection of toxoplasmone.

The present invention relates to an antibody specifically binding to Nectin-2 and uses thereof. A mouse anti-Nectin-2 antibody capable of specifically targeting Nectin-2 was discovered and analyzed for characteristics, and a chimeric anti-Nectin-2 antibody (chimeric 12G1; c12G1) was produced. In addition, in order to verify the specificity of mouse 12G1 antibody, siRNA for Nectin-2 was constructed and transduced into HEK-293 cells, and the expression level of Nectin -2 was confirmed through reverse transcription polymerase chain reaction, real-time polymerase chain reaction, and flow cytometry with fluorescent labeling. Furthermore, epitope mapping, enzyme-linked immunosorbent assay, surface plasmon resonance, fluorescence-activated cell sorting, and internalization assay were performed for characterization of the chimeric 12G1 antibody. Accordingly, the antibody developed in the present invention has specific reactivity to Nectin-2 and thus presents potential as an antibody therapeutic agent for cancer and autoimmune diseases.

A protein detection method and a detection device as well as use are provided. The protein detection method includes: fixing a probe type graphic encoding chip coupled with a specific capture antibody in the detection device, and meanwhile respectively sealing SAPE and a detection antibody in the detection device; and when in detection, performing detection and analysis based on an enzyme linked immunosorbent assay (ELISA) principle through the probe-type graphic encoding chip after the detection antibody and the SAPE are respectively dissolved, so as to obtain protein quantitative analysis results. In the protein detection method, the adopted Lab-in-Tip technology can complete processes such as sampling, washing, detection antibody hybridization, washing and binding of fluorescence labeled streptavidin and a biotin labeled detection antibody in such the Lab-in-Tip device, and has the advantages of multiplexed detection, such as quick completion, convenience, high flexibility, good specificity, few sample capacity and wide detection range.

This disclosure provides methods for identifying a subject suitable for an adeno associated virus (AAV) therapy. In some embodiments, the method comprises measuring a titer of an antibody or antigen-binding portion thereof that specifically binds to an AAV (“anti-AAV antibody”) in a biological sample obtained from the subject using an enzyme-linked immunosorbent assay (ELISA).

The application discloses a method for detecting tumor-specific T cells. After collecting whole cell components of tumor cells or tumor tissues, the free whole cell components or the whole cell lysate components loaded on nano / microparticles are incubated with peripheral immune cells. After the cancer-specific T cells are activated, the specific molecules of the tumor-specific T cells are detected to determine the content of the cancer-specific T cells in peripheral tissues such as peripheral blood. The whole cell lysate components are water-soluble components and non-water-soluble components, which are in a free state or loaded on nano / microparticles. The loading mode is that the water-soluble components and the non-water-soluble components of the whole cells are respectively or simultaneously loaded in the particle interior and / or respectively or simultaneously loaded on the particle surface. The detection method is flow cytometry, enzyme-linked immunospot technology, enzyme-linked immunosorbent assay technology, colloidal gold immunochromatography, gene detection technology and the like.

The invention discloses a recombinant protein of capsicum yellow ringspot virus, a polyclonal antibody and application of the recombinant protein and the polyclonal antibody, the recombinant protein of capsicum yellow ringspot virus is a CYRSV Nm recombinant protein, and the base sequence of the CYRSV Nm recombinant protein is as shown in SEQ ID No.1. Recombinant protein of the capsicum yellow ringspot virus N gene is used as an antigen to prepare a CYRSV Nm polyclonal antibody, an indirect-enzyme-linked immunosorbent assay (ID-ELISA) method and a spot-enzyme-linked immunosorbent assay (DOT-ELISA) method are used for detecting the capsicum yellow ringspot virus in a sample, and the method can realize rapid and high-throughput detection.

This invention relates to a prothioconazole hapten, artificial antigen, polyclonal antibody, and their preparation methods and applications. The structural formula of the prothioconazole hapten is shown in formula (1). When the prothioconazole hapten, artificial antigen, and polyclonal antibody of this invention are used for indirect competitive enzyme-linked immunosorbent assay (ELISA) to detect prothioconazole residues, the sample pretreatment is simple, the detection results are accurate, the detection time is short and the cost is low. At the same time, the operation is simple and samples can be processed in batches, which is conducive to the development of field work.

This invention belongs to the field of biomedical technology, specifically relating to a dissociation solution for aluminum adjuvant-adsorbed influenza vaccines and a method for determining antigen content. The dissociation solution of this invention is composed of potassium phosphate solution, diethanolamine, Triton X-100, Tween-20, dodecyl dimethyl betaine, and water. This dissociation solution exhibits excellent dissociation efficiency, achieving a recovery rate of 106%-110% for hemagglutinin from H1N1, H3N2, and BV influenza vaccines. It effectively protects antigen activity and is simple to operate with wide applicability. The dissociated sample can be directly used for mainstream detection methods such as one-way immunodiffusion assay, enzyme-linked immunosorbent assay, and high-performance liquid chromatography, enabling precise quantification of antigen content in aluminum adjuvant-adsorbed influenza vaccines and providing reliable technical support for vaccine quality control.

The invention discloses a double-nano-enzyme probe for detecting beta-amyloid oligomer, a kit based on the double-nano-enzyme probe as well as a preparation method and application of the double-nano-enzyme probe. The double nano-enzyme probe disclosed by the invention comprises a Si (at) Ag / Pt nano-enzyme probe functionalized by a beta-amyloid protein oligomer aptamer; the Ag / Pt nano-enzyme probe is hybridized and combined with the Si (at) Ag / Pt nano-enzyme probe functionalized by the beta-amyloid protein oligomer aptamer. The invention further discloses a preparation method of the double-nano-enzyme probe. The double-nano-enzyme probe disclosed by the invention can be used for constructing a nano-enzyme-linked immunosorbent assay (NLISA) platform with high sensitivity and high specificity, and accurate and quantitative detection of the beta-amyloid oligomer is realized through an antibody-antigen-aptamer sandwich method and a DNA cross-linking amplification strategy.

The present invention relates to a humanized antibody specifically binding to Nectin-2, and use thereof. The humanized anti-Nectin-2 antibody was prepared by humanizing a chimeric anti-Nectin-2 antibody capable of specifically targeting Nectin-2. The antigen binding equivalency of chimeric and humanized anti-Nectin-2 antibodies was identified using an enzyme-linked immunosorbent assay, and the affinity of the humanized anti-Nectin-2 antibody was measured. In addition, the Nectin-2 binding equivalency of the humanized anti-Nectin-2 antibody in live cells was identified, and the Nectin-2 binding equivalency expressed on the live cell surfaces of chimeric and humanized antibodies was also identified using flow cytometry. It was found that the humanized anti-Nectin-2 antibody of the present invention maintained freezing and thawing stability and also maintained pH-dependent Nectin-2 binding stability. Therefore, the humanized anti-Nectin-2 antibody developed in the present invention has specific reactivity to Nectin-2, and thus shows potential as an antibody therapeutic agent for cancer and autoimmune diseases.

The present invention discloses a camel-derived single-domain antibody GH12 that recognizes a conformational B cell antigen epitope in Clostridium difficile glutamate dehydrogenase and its application. The nucleotide sequence of the camel-derived single-domain antibody GH12 is shown in SEQ ID NO: 1, and the amino acid sequence is shown in SEQ ID NO: 2. The camel-derived single-domain antibody GH12 of the present invention has the advantages of strong specificity, high sensitivity, and good accuracy. It has been demonstrated that it can be used to detect recombinantly expressed CD-GDH and endogenous CD-GDH using methods such as enzyme-linked immunosorbent assay (ELISA) and protein dot blot hybridization (DB).

The idea of the patent includes a new principle, which is the detection of immune complexes using ELISA technology, which is a new and modern principle that is added to the four principles of ELISA, the new principle (the fifth ELISA) depends on detection of immune complexes, which consist of two different parts of proteins (an antigen - an antigen). This complex can be detected through this new principle, and it was applied to the immune complex (thyroid-stimulating hormone receptors and antibodies formed against it) Hashimoto's patients. It was considered an example of applying this theory, the immune complex was actually detected, its concentration determined, and compared to other techniques. A standard solution specially designed for this purpose was used to prove this theory. In this patent, a new kit is specifically designed to detect these immune complexes. Immune complex, the group is considered successful. In terms of use.

A method and corresponding kit for detecting Sjögren's Disease or predicting labial salivary gland biopsy results. The method includes evaluating an individual for the presence of one or more peptides or whole proteins or fragments thereof having an amino acid sequence selected from SEQ ID NOS: 1-67. The method and kit may be formatted as an enzyme-linked immunosorbent assay (ELISA).

Disclosed here is a device and method to detect the presence or absence of a target molecule by detecting fluorescence resonance energy transfer (FRET) signal generated when two labeled probes bind to the target molecule. The method is a sandwich enzyme-linked immunosorbent assay (ELISA) or a nucleic acid hybridization assay.

To provide a method for detecting an immunoglobulin in a sample, to provide the fusion protein to be used in this method, and to provide mutant luciferases with improved properties that notably can be used in this method.SOLUTION: A fusion protein includes an N-terminal domain which comprises an antibody which is a variable domain of a camelid heavy-chain antibody (VHH) or a single chain variable fragment (scFV) and which is directed against an immunoglobulin, and a C-terminal domain which comprises a polypeptide with a luciferase activity and which has a specific amino acid sequence or at least 80% amino acid sequence identity to the specific amino acid sequence.SELECTED DRAWING: None

The invention discloses a nano modification and high-density coating process for the surface of an enzyme-linked immunosorbent assay plate. The composition comprises an interface self-assembly bottom layer solution containing tannic acid and branched polyethyleneimine, and a dynamic gating functionalized buffer solution containing sodium tetraborate, D-mannitol and trehalose. The process comprises the following steps: firstly, constructing a supramolecular bottom layer on the surface of polystyrene, and then forming a boric acid-mannitol gating interface through non-washing drying; after the antibody is added, the Fc segment sugar chain and boric acid are subjected to competitive replacement reaction, so that mannitol is replaced, and the oriented high-density fixation of the antibody is realized. According to the invention, the problems of random antibody direction and weak binding force in traditional physical adsorption are solved, the antibody immobilization amount and detection sensitivity of the elisa plate are remarkably improved, and the elisa plate has good storage stability.

The invention discloses a nano antibody or an antigen binding fragment, a coding gene and a recombinant expression vector of a targeted porcine circovirus type 2 Rep protein, and application of the nano antibody or the antigen binding fragment, the coding gene and the recombinant expression vector as a reagent or a kit. The nanobody or the antigen binding fragment thereof comprises at least one complementary determinant selected from the group consisting of CDR1, CDR2 and CDR3. The high-affinity nano antibody molecule aiming at the porcine circovirus type 2 (PCV2) Rep protein, which is constructed by the invention, is combined with a competitive enzyme-linked immunosorbent assay system, so that high-sensitivity, high-specificity and high-stability quantitative detection on the anti-PCV2 antibody in porcine serum can be effectively realized. According to the technical scheme, multiple limitations of a traditional antibody detection method in the aspects of stability, production consistency, detection flux and cross reactivity are broken through, and a set of standardized diagnosis platform suitable for grass-roots breeding institutions, large-scale epidemiological screening and vaccine immune effect evaluation is constructed.

The application discloses a hybridoma cell strain secreting a monoclonal antibody against Xanthomonas oryzae pv. oryzae (Xoo) and application of the monoclonal antibody. A BALB / c mouse is immunized by using a cultured Xoo Y2 strain as an antigen, and a hybridoma cell strain 19C9 capable of secreting the monoclonal antibody against the Xoo is obtained through a hybridoma technology, and the strain is preserved with a preservation number of CGMCC No. 46004. The indirect ELISA titer of the monoclonal antibody secreted by the cell strain reaches 10 ‑7 , the antibody type and subclass are IgG1 and kappa light chain, the monoclonal antibody has specific immunoreactions with different strains or isolates of the Xoo, does not react with other bacteria such as rice bacterial leaf streak bacteria and rice panicle blast bacteria, and does not have an immunoreaction with healthy rice plant tissues. Dot-ELISA and immunocolloidal gold test strip two serological methods for detecting the Xoo are established by using the 19C9 monoclonal antibody. The hybridoma cell strain, the preparation of the monoclonal antibody secreted by the hybridoma cell strain and the establishment of the serological detection method provide detection reagents and technical support for detection and diagnosis, epidemiological investigation and scientific prevention and control of the Xoo.

The invention relates to a vaccine, in particular to a chimeric virus-like particle vaccine based on an echovirus 30 type key epitope as well as a preparation method and application of the chimeric virus-like particle vaccine. The invention provides a method for screening antigen epitopes, which comprises the following steps: firstly, predicting the antigen epitope of capsid protein VP1 through bioinformatics analysis, and then screening epitopes which can be effectively combined with anti-VP1 complete sequence serum through enzyme-linked immunosorbent assay. Results of an enzyme-linked immunosorbent assay show that the expressed antigen epitope peptide E30-6 (SEQ ID NO: 2) can be strongly combined with E30 VP1 antiserum, so that one high-immunogenicity antigen epitope peptide is screened based on the complete sequence of the echovirus 30 VP1, which has important guiding significance for research and development of echovirus 30 vaccines. Besides, the HBc-E30-6 vaccine is designed and prepared based on the epitope, the HBc-E30-6 vaccine is high in purity and can be self-assembled into VLP nanoparticles, and more importantly, serum immunized by the vaccine can effectively neutralize the echovirus 30.

The invention discloses a hepatitis A vaccine virus titer trace detection method, a kit and application. Comprising the following steps: inoculating human diploid cells into a porous cell culture plate, and culturing to form monolayer cells; inoculating a to-be-detected hepatitis A virus sample into the porous cell culture plate with monolayer cells, carrying out virus adsorption and culture, marking as an experimental group, and taking human diploid cells not inoculated with the virus sample as a negative control group; adding acetone into the experimental group and the negative control group for incubation so as to lyse cells and fix hepatitis A viruses and antigens thereof; detecting by adopting an enzyme-linked immunosorbent assay, and calculating according to a detection result to obtain the virus titer of the hepatitis A virus sample to be detected. The method is simple and convenient to operate, a virus extraction step in a traditional method is omitted, reagents are remarkably saved, the detection flux is improved, and the method is highly related to results of a pharmacopoeia method and is suitable for vaccine process research and development and quality control.