14 results about "Subcloning" patented technology

The invention belongs to the technical field of virus detection, and particularly relates to a porcine delta coronavirus N protein monoclonal antibody and a latex immunochromatography test strip containing the antibody. According to the invention, a prokaryotically expressed PDCoV N protein is used as an immunogen to immunize a mouse, and a specific monoclonal antibody 2G3 and a specific monoclonal antibody 9C4 are screened through cell fusion and a subcloning technology. The test strip comprises a PVC (polyvinyl chloride) bottom plate, a sample pad, a combination pad, a nitrocellulose membrane (NC membrane) and a water absorption pad, a quality control line C and a detection line T are respectively arranged on the NC membrane. The test strip can specifically recognize PDCoV, the lowest detection limit can reach 103.0 TCID50 / mL, the test strip has no cross reaction with other clinical common diarrhea viruses, the result interpretation only needs 15 min, and the test strip has the advantages of being rapid, sensitive, easy and convenient to operate and the like.

The application provides an anti-BDCA-2 antibody and a preparation method and application thereof, and belongs to the antibody field. The application provides a preparation method of an anti-BDCA-2 antibody, positive clones are screened by carrying out ELISA detection on hybridoma cell culture supernatant and antigen protein, and the anti-BDCA-2 antibody is prepared by subcloning the positive clones. The anti-BDCA-2 antibody comprises a heavy chain and a light chain, and the variable region of the heavy chain and the variable region of the light chain are selected from the amino acid sequences shown in SEQ ID NO:1-22. The anti-BDCA-2 antibody and related drugs provided by the application can be used for inhibiting pDC to generate IFN-alpha and preventing the excessive activation of pDC. The anti-BDCA-2 antibody generated by the application can effectively inhibit the activation of pDC cells and the generation of IFN-alpha, and has great potential in resisting virus infection and preventing certain autoimmune diseases.

The application provides a hybridoma cell strain secreting a monoclonal antibody against a alginate mannuronic acid tetrasaccharide epitope, a monoclonal antibody and application. In order to enhance the immunogenicity of the mannuronic acid tetrasaccharide, a glycoconjugate KLH-1 is used as an immunogen to inject and immunize a mouse, so that the mouse is stimulated to produce a specific immune response against the mannuronic acid tetrasaccharide epitope, then spleen cells of the immunized mouse are fused with myeloma cells, and after screening, an initial hybridoma cell strain is obtained; then after subcloning and specific screening, a hybridoma cell strain capable of stably secreting a monoclonal antibody of the target mannuronic acid tetrasaccharide epitope in alginate is obtained, and a monoclonal ascites antibody is further obtained. The monoclonal ascites antibody shows specific recognition and combination ability for pseudomonas aeruginosa, and the combination activity is related to the expression level of alginate on the surface of the bacteria. Therefore, the monoclonal ascites antibody can be used as a precise detection and diagnosis tool for pseudomonas aeruginosa, and can also be applied to the antibacterial treatment of pseudomonas aeruginosa infection.

This invention provides a monoclonal antibody against the VP2 protein of a mandarin fish two-segmented RNA virus, its preparation method, and its application. The VP2 protein was obtained by expression and purification using recombinant plasmids. Mice were immunized three times, and mice with the highest serum titer were selected for intraperitoneal booster immunization against VP2 protein. Mouse spleen cells and SP2 / 0 myeloma cells were fused, and hybridoma cell lines secreting VP2 protein antibodies were screened. These cells were then subcloned to obtain four stable hybridoma cell lines secreting anti-VP2 protein antibodies, and their heavy and light chain variable region sequences were determined. Mice were injected intraperitoneally with Freund's incomplete adjuvant, followed by the injection of the hybridoma cell lines. Ascites fluid was collected from the mice, extracted, purified, and used to obtain the monoclonal antibody against the VP2 protein. Using these four monoclonal antibodies, three highly conserved and specific antigenic epitopes of the VP2 protein of the mandarin fish two-segmented RNA virus were screened and identified. This invention provides support for establishing specific serological methods for mandarin fish two-segmented RNA viruses and for further research on the biological function and clinical diagnosis of VP2.

A blocking monoclonal antibody screening method based on a porcine PD-1 / PD-L1 monoclonal antibody and a heterologous Fc fusion protein comprises the following steps: (a) preparing a porcine PD-1 or PD-L1 extracellular region recombinant protein; (b) immunizing mice by using the recombinant protein to obtain immune spleen cells; (c) fusing the immune spleen cells with myeloma cells to obtain hybridoma cells; (d) screening and subcloning a hybridoma cell strain capable of secreting an anti-porcine PD-1 or PD-L1 antibody; (e) preparing a porcine PD-L1-rabbit Fc fusion protein; and (f) evaluating the blocking effect of the monoclonal antibody on PD-1 / PD-L1 binding by using the PD-L1-rabbit Fc fusion protein through flow cytometry.

The invention belongs to the technical field of biology, and particularly relates to a monoclonal antibody targeting African swine fever virus p30 protein and an epitope of the monoclonal antibody. A mouse is immunized by an African swine fever virus p30 recombinant protein, a cell strain is obtained by utilizing cell fusion and subcloning technologies, and a monoclonal antibody targeting the African swine fever virus p30 protein is generated, a heavy chain variable region of the monoclonal antibody comprises CDR1-3 shown as SEQ ID NO.1-3, and a light chain variable region of the monoclonal antibody comprises CDR1-3 shown as SEQ ID NO.4-6; the monoclonal antibody can specifically recognize and bind to the African swine fever virus, the amino acid sequence of the p30 protein antigen epitope recognized by the monoclonal antibody is shown as SEQ ID NO.11, and the epitope is highly conserved between African swine fever virus gene I type, African swine fever virus gene II type and African swine fever virus I / II type recombinant virus strains, and has important application value in the field of African swine fever virus research and detection analysis.

The invention provides an SP monoclonal antibody and a preparation method and application thereof. A heavy chain variable region nucleotide sequence and a light chain variable region nucleotide sequence of the SP monoclonal antibody are obtained by constructing a mouse bacteriophage library through screening and enrichment. Respectively subcloning the obtained heavy chain and light chain variable region nucleotide sequences into an expression vector containing an IgG1 constant region nucleotide sequence to construct a complete IgG1 antibody expression vector, and introducing the complete IgG1 antibody expression vector into eukaryotic cells for expression and purification to obtain an SP monoclonal antibody; and a monoclonal antibody with the strongest affinity and high specificity is screened out through ELISA (Enzyme-Linked Immunosorbent Assay) detection. Meanwhile, the screened SP monoclonal antibody and the screened CGRP monoclonal antibody are combined for use, so that the expression of macrophage interferon and the expression of inflammatory factors are improved, and the virus infection resistance of individuals is improved.

The present application relates to the technical field of biological medicine, and particularly relates to an anti-Aβ 42 Monoclonal antibody, preparation method and application. The present application adopts Aβ 42 protein of human origin as an immunogen to immunize BALB / c mice, extracts B lymphocytes of the spleen of the mice after successful immunization, fuses the B lymphocytes with mouse myeloma cells SP2 / 0 through cell fusion technology, and obtains hybridoma cell strains stably secreting anti-Aβ 42 monoclonal antibody after three rounds of subclone screening, so as to obtain anti-Aβ 42 monoclonal antibody; the anti-Aβ 42 monoclonal antibody can specifically combine with Aβ 42 protein, and can be used for immunological detection, has a broad market prospect, and can have important clinical significance for further development of biological monitoring technology and therapeutic drugs taking Aβ 42 as a target.

The invention belongs to the field of biological medicine, and particularly relates to an ANGPTL3 (anti-ANGPTL3) humanized monoclonal antibody stably transformed strain and application thereof. According to the application, an anti-ANGPTL3 humanized monoclonal antibody expression vector containing different nucleotide sequences is introduced into a host cell through electrotransfection, and a recombinant cell line with the highest yield is preliminarily screened out. The recombinant cell line is further screened for multiple times through main cloning and subcloning, and finally an ANGPTL3-resistant humanized monoclonal antibody cell strain with the advantages of high expression quantity, stability and controllability is screened out. The invention further discloses an ANGPTL3 humanized monoclonal antibody based on the cell, and the monoclonal antibody can be used for preparing a reagent for detecting ANGPTL3 and can also be used for preparing a medicine for preventing and / or treating kidney diseases or metabolism-related diseases.

The application is suitable for the technical field of tumor identification, and provides a tumor heterogeneity identification method, device, electronic equipment and storage medium. The application firstly locates tumor risk genes with consistency change, then identifies subclone specific genes related to tumor risk gene expression, determines subclone specific genes related to survival of a patient to a specified correlation degree, performs consistency clustering analysis on sample patients to obtain a classification label, constructs an optimal tumor prognosis model and screens an optimal image genome feature according to the tumor MRI image and the classification label of the sample patient, and finally analyzes an external tumor MRI image through the optimal tumor prognosis model and the optimal image genome feature, so that tumor heterogeneity quantitative analysis is realized on the premise that the patient is not caused trauma, the survival time of a tumor patient is predicted only through the tumor MRI image, and important theoretical basis and application value are provided for tumor precision medicine.

The invention relates to a monoclonal antibody capable of recognizing a human Hepatocyte antigen, a preparation method of the monoclonal antibody and application of the monoclonal antibody in immunodetection. According to the technical scheme, 628-731 loci, 813-882 loci and 628-731 loci are selected to be connected in series to form the antigen peptide, codon optimization is carried out, the antigen peptide becomes a gene segment suitable for being expressed in escherichia coli BL21, and finally the obtained recombinant protein contains a Hepatocyte protein segment and a histidine protein tag. The recombinant protein is used for immunizing a mouse, and through cell fusion, screening and subcloning, a mouse hybridoma cell strain capable of secreting the anti-Hepatocyte protein monoclonal antibody and the anti-Hepatocyte protein monoclonal antibody secreted by the cell strain are obtained. The antibody obtained by the scheme has high specificity and sensitivity, can specifically recognize cells expressing Hepatocyte protein, and is suitable for immunological detection, especially immunohistochemical detection.

The invention relates to a monoclonal antibody capable of recognizing a human MGMT antigen, a preparation method of the monoclonal antibody and application of the monoclonal antibody in immunodetection. According to the technical scheme, the full-length protein is selected as the antigen peptide, codon optimization is carried out, a gene segment suitable for being expressed in escherichia coli BL21 is formed, and the finally obtained recombinant protein contains an MGMT protein and a histidine protein tag. The recombinant protein is used for immunizing a mouse, and through cell fusion, screening and subcloning, a mouse hybridoma cell strain secreting the anti-MGMT protein monoclonal antibody and the anti-MGMT protein monoclonal antibody secreted by the cell strain are obtained. The antibody obtained by the scheme has high specificity and sensitivity, can specifically recognize cells expressing MGMT protein, and is suitable for immunological detection, especially immunohistochemical detection.

The present application relates to a kind of monoclonal antibody that can identify human PSA antigen, secreting cell strain, its preparation method and its use in immunodetection.The above technical scheme selects the amino acid of 31-124 of PSA protein as antigen peptide, is codon optimized, becomes the gene fragment suitable for expression in escherichia coli BL21, and finally the recombinant protein includes TRX protein label, PSA protein fragment and histidine protein label.The recombinant protein is immunized to mouse, is fused by cell, screening and subcloning, obtains the mouse hybridoma cell strain 16F1 of high-efficiency secreted anti-PSA protein monoclonal antibody and the anti-PSA protein monoclonal antibody secreted by the cell strain.The antibody obtained in the present application has high specificity, sensitivity, and can specifically recognize the cell expressing PSA protein, is suitable for immunological detection, especially immunohistochemical detection.

This invention belongs to the field of virus detection technology, specifically relating to a monoclonal antibody against porcine delta coronavirus N protein and a latex immunochromatographic test strip containing this antibody. This invention uses prokaryotically expressed PDCoV N protein as an immunogen to immunize mice, and screens for specific monoclonal antibodies 2G3 and 9C4 through cell fusion and subcloning techniques. The test strip includes a PVC base plate, a sample pad, a conjugation pad, a nitrocellulose membrane (NC membrane), and an absorbent pad; a control line C and a detection line T are respectively set on the NC membrane. This test strip can specifically recognize PDCoV, with a detection limit as low as 10. 3.0 TCID 50 The virus has a concentration of 1000 μL / mL and does not cross-react with other common clinical diarrhea viruses. The results can be interpreted in just 15 minutes, making it fast, sensitive, and easy to operate.