61 results about "Subcloning" patented technology

The invention belongs to the technical field of biology, and particularly relates to a latex duplex immunochromatography test strip for rapidly identifying a porcine transmissible gastroenteritis virus (TGEV) and a porcine epidemic diarrhea virus (PEDV) and a preparation method of the latex duplex immunochromatography test strip. Soluble TGEV N protein and PEDV N protein with immunogenicity and reactivity are respectively obtained by utilizing a prokaryotic expression system, the soluble TGEV N protein and the soluble PEDV N protein are used as immunogenicity to immunize mice, and specific monoclonal antibodies of the TGEV N protein and the PEDV N protein are respectively obtained by utilizing cell fusion and subcloning screening technologies. The antibody variable region gene sequence is obtained by using a nested PCR amplification technology, and can be subsequently obtained by using a gene engineering or protein engineering method. Then, the latex duplex immunochromatography test strip for detecting TGEV and PEDV, which is convenient, rapid and high in sensitivity, and the preparation method are researched and developed on the basis of specific monoclonal antibodies of TGEV N protein and PEDV N protein.

The invention discloses a herbicide-resistant protein GAT hybridoma cell strain, an antibody generated by the herbicide-resistant protein GAT hybridoma cell strain and an application of the herbicide-resistant protein GAT hybridoma cell strain. The herbicide-resistant protein GAT hybridoma cell strain is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.46333. The preparation method comprises the following steps: a) purifying prokaryotic expression to obtain GAT recombinant protein; b) immunizing animals: immunizing BALB / c mice by taking the GAT recombinant protein as an antigen; c) cell fusion: splenocytes of the immune BALB / c mice are collected and fused with SP2 / 0 cells; and d) cell strain establishment: carrying out subcloning through a limited dilution method, carrying out ELISA detection after 5-7 days of subcloning until a hybridoma cell strain capable of stably secreting a positive antibody is screened out, and carrying out amplification, re-culture and storage. The monoclonal antibody secreted by the hybridoma cell strain lays a foundation for detection of herbicide-resistant protein GAT in transgenic crops, so that development, seed production and seed conservation of transgenic gat crops are supported, and stable industrial promotion of the transgenic gat crops and sustainable application of related transformants are guaranteed.

An anti-BDCA-2 antibody, a preparation method therefor, and a use thereof, relating to the field of antibodies. The preparation method for the anti-BDCA-2 antibody comprises: carrying out ELISA detection and screening on a hybridoma cell culture supernatant and an antigen protein to obtain a positive clone, and subcloning the positive clone to obtain the anti-BDCA-2 antibody. The anti-BDCA-2 antibody comprises a heavy chain and a light chain, wherein a variable region of the heavy chain and a variable region of the light chain are selected from amino acid sequences as shown in SEQ ID NOs: 1-22. The anti-BDCA-2 antibody and related drugs can be used for inhibiting the production of IFN-α by pDCs and preventing the over-activation of the pDCs. The anti-BDCA-2 antibody can effectively inhibit the activation of pDC cells and the production of IFN-α, and has great potential in resisting virus infection and preventing certain autoimmune diseases.

Disclosed herein is Differential Subclone Eradication and Resistance Analysis (DSER), a method developed to identify molecular targets for improved therapy by direct comparison of genomic features of eradicated and resistant subclones in pre-and post-treatment samples from a patient with BRCA2-deficient metastatic prostate cancer. FANCI and EYA4 were identified as candidate DNA repair-related targets for converting subclones from resistant to eradicable, and RNAi-mediated depletion of FANCI confirmed it as a potential target. The EYA4 alteration was associated with adjacent L1 transposon insertion during cancer evolution upon treatment. L1 activation was inhibited by the antiretroviral drug azidothymidine. In conclusion DSER provides an informative intermediate step toward effective precision cancer medicine, especially in cases with dramatic but temporary metastatic tumor regression. L1 transposon activation may be a modifiable source of cancer genomic heterogeneity, suggesting the potential of leveraging newly discovered triggers and blockers of L1 activity to overcome therapy resistance.

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

The invention discloses a herbicide-resistant protein GAT hybridoma cell strain, an antibody generated by the herbicide-resistant protein GAT hybridoma cell strain and application of the herbicide-resistant protein GAT hybridoma cell strain and the antibody. The herbicide-resistant protein GAT hybridoma cell strain is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.46332. The preparation method comprises the following steps: a) purifying prokaryotic expression to obtain GAT recombinant protein; b) immunizing animals: immunizing BALB / c mice by taking the GAT recombinant protein as an antigen; c) cell fusion: splenocytes of the immune BALB / c mice are collected and fused with SP2 / 0 cells; and d) cell strain establishment: carrying out subcloning through a limited dilution method, carrying out ELISA detection after 5-7 days of subcloning until a hybridoma cell strain capable of stably secreting a positive antibody is screened out, and carrying out amplification, re-culture and storage. The monoclonal antibody secreted by the hybridoma cell strain lays a foundation for detection of herbicide-resistant protein GAT in transgenic crops, so that development, seed production and seed conservation of transgenic gat crops are supported, and stable industrial promotion of the transgenic gat crops and sustainable application of related transformants are guaranteed.

The invention relates to the technical field of gene engineering, in particular to application of a SmOPR3 gene in creation of a male sterile line of salvia miltiorrhiza. The invention provides application of a SmOPR3 gene in creation of a male sterile line of salvia miltiorrhiza. The nucleotide sequence of the SmOPR3 gene is shown as SEQ ID NO: 1. Experimental results show that three regenerated salvia miltiorrhiza bunge are obtained in this conversion, and results of PCR amplification and Sanger sequencing on a SmOPR3 mutation region show that the three salvia miltiorrhiza bunge strains are effectively edited. The subcloning analysis confirms that the three materials successfully realize accurate and synchronous editing of two targets, the editing form of one salvia miltiorrhiza (no.1) realizes allele knockout, and the deletion of SmOPR3 directly leads to maldevelopment of stamens of salvia miltiorrhiza, cannot normally generate pollen and finally leads to the phenotype of male sterility.

The application provides a kind of MFGE8 monoclonal antibody and its preparation method and application, the heavy chain and light chain variable region nucleotide sequence of the MFGE8 monoclonal antibody is obtained by constructing rabbit phage library, panning, monoclonal screening and identification, then the heavy chain and light chain variable region nucleotide sequence obtained is respectively subcloned into the expression vector containing IgG constant region nucleotide sequence, is constructed into complete IgG antibody expression vector and is introduced into eukaryotic cell to express and purify, and five strains of monoclonal antibodies with strong affinity are screened out by ELISA experiment.The application also pairs five strains of monoclonal antibodies two by two to screen the antibody pair with the strongest antigen binding capacity, and uses the antibody pair to prepare MFGE8 ELISA detection kit for quantitatively detecting the content of MFGE8 in human biological samples, to assist early diagnosis and screening of breast cancer.

The present application relates to a kind of monoclonal antibody that can identify human Topo II alpha antigen, secretes cell strain, its preparation method and its use in immunodetection.The above technical solution selects the amino acid of 1301-1531 of Topo II alpha protein as antigen peptide, codon optimization becomes the gene fragment suitable for expression in escherichia coli BL21, and finally the recombinant protein includes Topo II alpha protein fragment and histidine protein label.The recombinant protein is immunized to mouse, is fused by cell, screening and subcloning, and obtains mouse hybridoma cell strain, and the anti Topo II alpha protein monoclonal antibody secreted by the cell strain is obtained.The antibody obtained by the present application has high specificity, sensitivity, and can specifically recognize the cell expressing Topo II alpha protein, and is suitable for immunological detection, especially immunohistochemical detection.

The invention provides a PEDV (porcine epidemic diarrhea virus) N protein monoclonal antibody as well as a preparation method and application thereof, and the method comprises the following steps: performing prokaryotic expression on PEDV N protein, and purifying to obtain the PEDV N protein; emulsifying the purified PEDV N protein and an isopyknic Freund's adjuvant, injecting the emulsified PEDV N protein into 8-week-old female BALB / c mice at multiple points, and immunizing once every two weeks; the method comprises the following steps: uniformly mixing splenocytes of a mouse subjected to boosted immunization for 3 days with SP2 / 0 cells in a logarithmic phase according to a ratio of 5: 1, carrying out cell fusion under the action of PEG 1500, and culturing a fused cell mixed solution by using an HAT culture medium; screening out positive pores with strong antibody secretion capacity by adopting an indirect ELISA (Enzyme-Linked Immunosorbent Assay) method, and carrying out subcloning by adopting a limited dilution method to obtain six cell strains for secreting antibodies; the PEDV N protein mAb is prepared by adopting an in-vivo ascites inducing method. The prepared PEDV N protein monoclonal antibody can be specifically and efficiently combined, and can be used as an antibody tool for researching PEDV virus infection and anti-infection mechanisms.

The invention relates to a photo-enzyme catalysis system based on sulfur metabolism of yarrowia lipolytica and application of the photo-enzyme catalysis system, and belongs to the technical field of genetic engineering and microbial fermentation. The photo-enzyme catalysis system for sulfur metabolism of Yarrowia lipolytica comprises Yarrowia lipolytica, a cadmium binding polypeptide biosynthesis module and a ferredoxin NADP (+) reductase biosynthesis module, the cadmium binding polypeptide biosynthesis module and the ferredoxin NADP (+) reductase biosynthesis module are sequentially connected through subcloning, and the cadmium binding polypeptide biosynthesis module and the ferredoxin NADP (+) reductase biosynthesis module are sequentially connected through subcloning. The yeast cells are located on Yarrowia lipolytica cell chromosomes; the Met25 gene in the recombinant yarrowia lipolytica is knocked out. A light capture and electron transfer system is constructed by recombining a cadmium binding polypeptide gene sequence and a ferredoxin NADP (+) reductase gene in yeast cells, and the oil yield of yarrowia lipolytica is increased.

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 present application relates to a kind of monoclonal antibody that can identify human smooth muscle myosin antigen, preparation method and its use in immunodetection.The above technical solution selects the amino acid of 1180-1400 of smooth muscle myosin as antigen peptide, codon optimization becomes the gene fragment suitable for expression in escherichia coli, and finally the recombinant protein includes smooth muscle myosin fragment and histidine protein label.The recombinant protein is immunized to mouse, and is subjected to cell fusion, screening and subcloning, obtains mouse hybridoma cell strain 15C1D4 and the anti-smooth muscle myosin monoclonal antibody secreted by the cell strain.The antibody obtained by the present application has high specificity, sensitivity, and can specifically recognize the cell expressing smooth muscle myosin, and is suitable for immunological detection, especially immunohistochemical detection.

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.

The application provides a CD93 monoclonal antibody and a preparation method and application thereof, the heavy chain and light chain variable region nucleotide sequences of the CD93 monoclonal antibody are obtained by screening and enrichment through constructing a rabbit phage library, then the obtained heavy chain and light chain variable region nucleotide sequences are respectively subcloned into an expression vector containing an IgG1 constant region nucleotide sequence, an entire IgG1 antibody expression vector is constructed and introduced into a eukaryotic cell for expression and purification, and the CD93 monoclonal antibody is obtained; three strains of CD93 monoclonal antibodies with the strongest affinity are screened out through ELISA detection. The application further pairs the three strains of screened monoclonal antibodies two by two to screen out an antibody pair with strong antigen binding capacity, and prepares a CD93 ELISA kit from the antibody pair and uses the antibody pair for quantitative detection of the CD93 content in a human biological sample, thereby assisting in the diagnosis and prognosis evaluation of hepatocellular carcinoma.

The application relates to a kind of monoclonal antibody that can identify human Myogenin antigen, its preparation method and its use in immunodetection.The above technical solution selects the amino acid of 1-224 of Myogenin protein as antigen peptide, is codon optimized, becomes the gene fragment suitable for expressing in escherichia coli BL21, and finally the recombinant protein contains Myogenin protein fragment and histidine protein label.The recombinant protein is immunized to mouse, is fused to cell, is screened and subcloned, obtains the mouse hybridoma cell strain of secreting anti-Myogenin protein monoclonal antibody and the anti-Myogenin protein monoclonal antibody secreted by the cell strain.The antibody obtained in the scheme has high specificity, sensitivity, can specifically recognize the cell expressing Myogenin protein, is suitable for immunological detection, especially immunohistochemical detection.

The application belongs to the technical field of biology and particularly relates to a latex double immunochromatographic test strip for rapidly identifying porcine transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhea virus (PEDV) and a preparation method thereof. The soluble TGEV N protein and PEDV N protein with immunogenicity and reactivity are obtained by using a prokaryotic expression system, mice are immunized by using the immunogenicity, and the specific TGEV N protein and PEDV N protein monoclonal antibodies are obtained by using cell fusion and subcloning screening technology. The antibody variable region gene sequence is obtained by using nested PCR amplification technology and can be obtained by using genetic engineering or protein engineering method subsequently. Then, the latex double immunochromatographic test strip for detecting TGEV and PEDV and the preparation method thereof are developed based on the specific TGEV N protein and PEDV N protein monoclonal antibodies.

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.

The present disclosure provides methods for predicting whether a patient diagnosed with cancer will benefit from treatment with an immune checkpoint inhibitor based on subclonal mutation levels.

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 discloses a hybridoma cell strain based on type 1 goose astrovirus, a construction method, a monoclonal antibody and a detection reagent. The hybridoma cell strain is a hybridoma cell strain A5A1, the preservation number of the hybridoma cell strain is CCTCC (China Center for Type Culture Collection) NO: C202505, and the hybridoma cell strain is preserved in the China Center for Type Culture Collection on March 26, 2025. According to the invention, a mouse is immunized by GAstV-1ORF2 recombinant protein expressed by a prokaryotic expression system, and a monoclonal antibody A5A1 is obtained by screening and subcloning. Indirect competitive ELISA (icELISA) for detecting the GAstV-1 antibody is developed based on the hybridoma cell strain A5A1, and the method is high in sensitivity, good in repeatability and high in specificity, only reacts with serum of GAstV-1, and does not have cross reaction with other detected pathogenic serum. A serological detection method established by the monoclonal antibody prepared by the method lays a foundation for scientific prevention and control of gout of goslings.

The invention provides a CD93 monoclonal antibody as well as a preparation method and application thereof. Heavy chain and light chain variable region nucleotide sequences of the CD93 monoclonal antibody are obtained by constructing a rabbit phage 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 a CD93 monoclonal antibody; three CD93 monoclonal antibodies with the strongest affinity are screened out through ELISA (enzyme-linked immuno sorbent assay) detection. According to the invention, the three screened monoclonal antibodies are pairwise paired to screen antibody pairs with strong antigen binding capacity, and the antibody pairs are used for preparing a CD93 ELISA kit which is used for quantitatively detecting the content of CD93 in a human biological sample and assisting in diagnosis and prognosis evaluation of hepatocellular carcinoma.

The present application relates to a kind of monoclonal antibody that can identify human Mucin6 antigen, its preparation method and its use in immunodetection.The above technical solution selects the amino acid of 1950-2050 of Mucin6 protein as antigen peptide, codon optimization becomes the gene fragment suitable for expressing in escherichia coli BL21, and finally the recombinant protein containing Mucin6 protein fragment and histidine tag.The recombinant protein is immunized to mouse, and after cell fusion, screening and subcloning, obtain the mouse hybridoma cell strain 37D10 and the anti-Mucin6 protein monoclonal antibody secreted by the cell strain.The antibody obtained by the present application has high specificity, sensitivity, and can specifically recognize the cell expressing Mucin6 protein, and is suitable for immunological detection, especially immunohistochemical detection.

In an aspect, there is provided a method for diagnosing or prognosing a subject with cancer, the method comprising: providing cancer DNA sequencing data from a cancer sample comprising cancer DNA from the subject; comparing the cancer DNA sequencing data with control DNA sequencing data to determine genetic aberrations; determining, from the genetic aberrations, the clonal and subclonal populations present in the sample; constructing a phylogenetic map of the clonal and subclonal populations; assigning to the subject a risk level associated with a better or worse patient outcome or response to therapy; wherein a relatively higher risk level is associated with a higher level of evolution and number of subclonal populations and a relatively lower risk level is associated with a lower level of evolution and number of subclonal populations.

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 discloses an IGHV region gene locus assembly based on Gibson assembly as well as a method and application of the IGHV region gene locus assembly. The assembly method comprises the following steps: designing upstream and downstream specific PCR (Polymerase Chain Reaction) primers for amplifying each target gene segment to obtain a corresponding single target gene segment, transforming a universal vector pUC19 into a subcloning vector, and obtaining a corresponding pMIGH-IGHV plasmid in a Gibson assembly mode; the preparation method comprises the following steps: carrying out Gibson assembly and double enzyme digestion, and then carrying out Gibson assembly to obtain pCC1BAC-IGHV (1-6V), pCC1BAC-IGHV (1-10V) and pCC1BAC-IGHV (1-15V), so as to obtain an assembled product. According to the assembling method disclosed by the invention, a plurality of target gene segments as long as 3.5 kb can be sequentially assembled according to a design sequence, and an assembled product can reach a hundred kb level or even an Mb level. According to the assembly mode provided by the invention, the human IGHV region gene locus can be simply and effectively prepared, and the human IGHV region gene locus can be applied to preparation of diversified antibodies according to conventional requirements.

The invention relates to the technical field of bioengineering, in particular to a hybridoma cell strain, a goat pox virus G9 protein monoclonal antibody and application. The hybridoma cell strain is named as 9C5, and is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.46312. Goat pox virus G9 protein immunogen is prepared, hybridoma cells capable of stably secreting antibodies are prepared through cell fusion and multi-time subcloning, a large number of goat pox virus G9 protein monoclonal antibodies are obtained through ascites preparation, the goat pox virus G9 protein monoclonal antibodies can still be effectively combined with goat pox viruses after being diluted by 64K times, and the goat pox virus G9 protein monoclonal antibodies can be used for detecting the goat pox viruses. The method can be used for constructing a goat pox virus detection method and lays a foundation for prevention and treatment of goat pox viruses.

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.