31 results about "Subcloning" patented technology

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 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 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.

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 application discloses a chicken salmonella pullorum strain stably expressing variant O antigen and a construction method and application thereof, and constructs an expression plasmid containing a J23119 promoter and a SPUL_2404 gene, and then the expression plasmid is transferred into chicken salmonella pullorum, so that a chicken salmonella pullorum strain overexpressing the SPUL_2404 gene is constructed to stably express the variant O antigen, and agglutination test shows that the strain only has a typical agglutination reaction with chicken salmonella pullorum variant serums, and does not have a typical agglutination reaction with chicken salmonella pullorum standard serums, and after subculture, O antigens of all subclone colonies are stable, and only have a typical agglutination reaction with the variant serums, so that the strain can be used for preparing chicken salmonella pullorum variant agglutination antigens.

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 discloses a herbicide-resistant protein GAT hybridoma cell strain, an antibody produced by the hybridoma cell strain and application of the hybridoma cell strain, and the hybridoma cell strain is preserved in the China General Microbiological Culture Collection Center with a preservation number of CGMCC No.46333.A preparation method of the hybridoma cell strain comprises the following steps: a) purifying a GAT recombinant protein obtained by prokaryotic expression; b) immunizing animals: using the GAT recombinant protein as an antigen to immunize BALB / c mice; c) cell fusion: collecting spleen cells of the immunized BALB / c mice and fusing the spleen cells with SP2 / 0 cells; d) cell strain establishment: performing subcloning by a limited dilution method, performing ELISA detection on the subcloning for 5-7 days, and screening the hybridoma cell strain stably secreting positive antibodies until the hybridoma cell strain is expanded, cultured and preserved again.The monoclonal antibody secreted by the hybridoma cell strain lays a foundation for realizing detection of the herbicide-resistant protein GAT in a transgenic crop, thereby supporting the development, seed production, seed preservation and guarantee of the transgenic crop, and stably promoting industrialization of the transgenic crop and sustainable application of related transformants. gat The application discloses a herbicide-resistant protein GAT hybridoma cell strain, an antibody produced by the hybridoma cell strain and application of the hybridoma cell strain, and the hybridoma cell strain is preserved in the China General Microbiological Culture Collection Center with a preservation number of CGMCC No.46333.A preparation method of the hybridoma cell strain comprises the following steps: a) purifying a GAT recombinant protein obtained by prokaryotic expression; b) immunizing animals: using the GAT recombinant protein as an antigen to immunize BALB / c mice; c) cell fusion: collecting spleen cells of the immunized BALB / c mice and fusing the spleen cells with SP2 / 0 cells; d) cell strain establishment: performing subcloning by a limited dilution method, performing ELISA detection on the subcloning for 5-7 days, and screening the hybridoma cell strain stably secreting positive antibodies until the hybridoma cell strain is expanded, cultured and preserved again.The monoclonal antibody secreted by the hybridoma cell strain lays a foundation for realizing detection of the herbicide-resistant protein GAT in a transgenic crop, thereby supporting the development, seed production, seed preservation and guarantee of the transgenic crop, and stably promoting industrialization of the transgenic crop and sustainable application of related transformants. gat The application discloses a herbicide-resistant protein GAT hybridoma cell

This invention discloses a hybridoma cell line containing the herbicide-resistant protein GAT, its produced antibody, and its applications. The hybridoma cell line is deposited at the China General Microbiological Culture Collection Center (CGMCC), accession number CGMCC No. 46332. The preparation method includes: a) purifying prokaryotic expression to obtain recombinant GAT protein; b) immunizing animals: using the recombinant GAT protein as an antigen to immunize BALB / c mice; c) cell fusion: collecting spleen cells from immunized BALB / c mice and fusing them with SP2 / 0 cells; d) cell line establishment: subcloning using the limiting dilution method, performing ELISA detection on subclones after 5-7 days, until a stable hybridoma cell line secreting positive antibodies is selected for expansion, reculturing, and preservation. The monoclonal antibody secreted by the hybridoma cell line of this invention lays the foundation for the detection of the herbicide-resistant protein GAT in transgenic crops, thus supporting the development of transgenic crops. street The development, seed production, and preservation of genetically modified crops have ensured the survival of genetically modified organisms (GMOs). street The industrialization of genetically modified crops is progressing steadily, and the sustainable application of related transformants is advancing.

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.

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 discloses an application of limonene in preparation of a reagent for inhibiting the heterogeneity drug resistance of staphylococcus aureus. The method comprises the following steps: firstly, respectively determining the minimum inhibitory concentrations of limonene and ciprofloxacin to a clinical ciprofloxacin heterogeneity drug-resistant staphylococcus aureus original bacterium and drug-resistant subcloning by adopting a trace broth dilution method, and designing a chessboard method drug concentration range on the basis of the minimum inhibitory concentrations; the result of a chessboard method shows that when the ciprofloxacin is combined with the limonene with the concentration of 640 mu g / mL, the growth of the drug-resistant subcloning of the heterogeneity drug-resistant staphylococcus aureus of the ciprofloxacin can be effectively inhibited, and good antibacterial activity is shown. In addition, a bacterial flora spectral pattern analysis experiment result shows that the limonene can effectively reduce the drug-resistant subgroup frequency under the condition of the concentration of 640 mu g / mL.

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.

Via yeast display technology, albumin binding variants were selected, and variants displayed improved binding to MSA or both HSA & MSA. All variants were subcloned into the pHEN-expression plasmid and expressed in E coli WK6 cells. Purified nanobodies were then analyzed for their capacity to bind immobilized MSA. Biotinylated nanobodies were then tested in vivo for their circulatory half-life. They then validated the effect of their nanobodies anti albumin linked with another nanobodies such as single-domain antibody anti-D′D3 as described in WO2017129630 and WO2018091621.Mice having reduced levels of VWF (Von Willebrand disease type-1) received a single dose of the bispecific single-domain antibody. VWF and FVIII levels were followed over 14 days. These data show that there is a statistically significant increase in VWF and FVIII levels, which is maintained for 10 days.The present invention relates to an isolated single-domain antibody (sdAb) directed against albumin and its uses in different diseases such as bleeding disorders.

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 invention discloses lactococcus lactis for promoting intestinal colonization. The lactococcus lactis is recombinant lactococcus lactis capable of expressing flagellin. Accordingly, a corresponding construction method is also established, that is, the flagellin gene is subcloned to a lactococcus lactis expression vector, and then recombinant plasmids are transformed into lactococcus lactis to obtain the lactococcus lactis. Research shows that the recombinant lactococcus lactis with the mFliC3 and the rFliC3 shows an adhesion promoting function in a cell co-culture experiment, and in a sterile mouse experiment, although the bacteria discharge time of the mFliC3 and the rFliC3 is longer than that of an empty vector group, the bacteria discharge time of the rFliC3 is longer than that of the mFliC3. Therefore, both the mFliC3 and the rFliC3 have the function of promoting field planting, and the field planting time of the rFliC3 is longer. Therefore, the intestinal colonization promoting lactococcus lactis disclosed by the invention can be used as probiotics or functional bacteria, is particularly applied to related food or health care products, and can be used for reducing the supplementing times or prolonging the supplementing interval time.

The present application relates to a kind of monoclonal antibody that can identify human Pit-1 antigen, its preparation method and its use in immunodetection.The above technical scheme synthesizes the amino acid of 372-448 of Pit-1 protein as antigen peptide, obtains immunogen after coupling KLH.The Pit-1-KLH protein is immunized to mouse, is fused to cell, screening and subcloning, obtain mouse hybridoma cell strain 4G5, and the monoclonal antibody secreted by the cell strain anti Pit-1 protein.The antibody obtained in the present application has high specificity, sensitivity, can specifically recognize the cell expressing Pit-1 protein, is suitable for immunological detection, especially immunohistochemical detection.

Methods and systems for analyzing genetic variants are disclosed. The methods and systems can be used to classify a variant in a tumor sample as germline, somatic, subclonal, or ambiguous. The variant can be classified by fitting a genome-wide copy number model to a sequence coverage input and a SNP allele frequency input. The fitted model can be used to determine a tumor purity, and a total copy number and a minor allele copy number for each of a plurality of genomic segments. Using the tumor purity, the total copy number and the minor allele copy number for the genomic segment comprising the variant, and the variant allele frequency for the variant in the tumor sample, the variant can be classified as germline, somatic, subclonal or ambiguous.

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.