84 results about "293t cell" patented technology

The invention relates to a CHO-S cell strain capable of stably expressing H5N1 hemagglutinin protein and a construction method of the CHO-S cell strain, and belongs to the field of bioengineering.H5N1 hemagglutinin protein expression plasmids are obtained in an in-vitro synthesis and seamless cloning mode, 293T cells are transfected through the H5N1 hemagglutinin protein expression plasmids, and it is proved that the H5N1 hemagglutinin protein is expressed; the CHO-S cell strain capable of stably expressing the H5N1 hemagglutinin protein is obtained by transfecting CHO-S cells by using the H5N1 hemagglutinin protein expression plasmids and carrying out multiple rounds of cloning and screening, so that a basis is provided for obtaining H5N1 recombinant protein influenza vaccines.

The invention discloses a method for quickly rescuing bovine coronavirus epidemic strains and application of the method. The bovine coronavirus SHZ isolate is subjected to efficient segmented cloning, the obtained segment and a Linker sequence are subjected to a cyclic polymerase extension reaction, and the obtained reaction product can be directly transfected to 293T cells for virus packaging without purification. Different from a traditional method, the method does not need to amplify full-length virus cDNA clone by means of bacteria and yeast, but directly obtains a sufficient amount of preliminary products through PCR, and avoids the problems of instability and low efficiency when partial sequences of a virus genome proliferate in a bacteria or yeast host. By adopting the reverse genetic system, the recombinant bovine coronavirus expressing the foreign protein is successfully rescued, a brand new technical platform is provided for dynamic visualization research of in-vivo and in-vitro replication of the virus, and an efficient and flexible tool is also provided for virology research, development of virus vector vaccines and screening of antiviral drugs.

The invention provides brain cell specific targeting delivery based on exosome, and the exosome secreted by 293T cells is used for engineering transformation, so that the capabilities of penetrating BBB and specifically targeting neurons are achieved. According to the present invention, the L1CAM is loaded on the exosome, the exosome is obtained by transfecting a 293T cell of a Lamp2b-L1CAM PCDH-HygroHA plasmid through a 293T-Lamp2b-L1CAM cell line, and the BBB cell-penetrating peptide 4F-T7 and the neuronal targeting protein L1CAM are loaded on the surface of the exosome so as to treat the neurodegenerative diseases such as the Alzheimer's disease and the Parkinson's disease.

The invention discloses a polyketone compound derived from actinomycetes as well as a preparation method and application thereof, and relates to the technical field of microbial natural product mining and biological medicine, and the key point of the technical scheme is that the invention discloses a novel polyketone compound Strepactone derived from actinomycetes Streptomyces sp. DP0001, and the molecular formula of the novel polyketone compound Strepactone is C25H38O5. The compound is obtained through strain fermentation, ethyl acetate extraction and multi-step chromatographic purification, and the structure is identified through ESI-MS, 1H NMR and 13C NMR. Experiments show that Strepactone has an inhibition effect on staphylococcus aureus and methicillin-resistant staphylococcus aureus and has remarkable inhibition activity on human non-small cell lung cancer A549 cells, the half inhibitory concentration IC50 of the Strepactone is 5.36 mu M, and the Strepactone has no obvious cytotoxic activity on human embryo kidney cells 293T cells at the concentration of 30 mu M. The compound provided by the invention can be used for preparing anti-drug-resistant bacteria drugs and anti-human non-small cell lung cancer drugs, and also provides a structural basis for the development of novel antibacterial and anti-tumor leading drugs.

The invention discloses a reverse transcriptase, wherein a coding sequence of the reverse transcriptase is derived from a Rattus norvegicus genome. The protein sequence of the protein comprises a protein sequence of a wild type Rattus norveicus, or a protein sequence which is subjected to engineering modification on the basis of the wild type Rattus norveicus, or a protein sequence which is subjected to engineering modification on the basis of the wild type Rattus norveicus. Furthermore, the protein sequence of the gene is as shown in any one of SEQ No.1-55. In a rice genome site and a human HEK 293T cell genome site, the system shows efficient editing efficiency of a guide editor. The invention provides a series of efficient reverse transcriptase components which can be carried on a guide editing system, important bottom-layer technical support is provided for research of point mutation in a genome, creation of a special site disease model and correction of genetic mutation sites, and the reverse transcriptase component has a wide application prospect in the field of precision medicine and precision agriculture breeding.

The present application relates to the technical field of genetic engineering, in particular to a T7-293T cell and application of the T7-293T cell in bovine parainfluenza virus type 3 infectious clone rescued virus, and the T7-293T cell provided by the present application has the preservation number of CGMCC No.46335.The T7-293T cell provided by the present application is a 293T cell stably expressing T7 RNA polymerase, and has the advantages of high transfection efficiency and cheap transfection reagent.Further, the present application establishes a method for bovine parainfluenza virus type 3 infectious clone rescued virus based on the T7-293T cell, constructs three genes of NP, P and L of bovine parainfluenza virus type 3 into the same helper plasmid, and co-transfects the T7-293T cell with the full-length gene plasmid of the virus, so as to avoid the low efficiency of co-transfecting four plasmids, and further improve the success rate of bovine parainfluenza virus type 3 infectious clone rescued virus.

The invention belongs to the technical field of exosomes, and particularly relates to an engineered exosome for overexpressing EGF and FGF as well as a preparation method, a product and application of the engineered exosome. An LAMP2B fusion expression technology is adopted, EGF and FGF are directionally anchored to the surface of a 293T cell source exosome membrane, free factor degradation is avoided, and the problem that the carrying amount of natural exosome factors is insufficient is solved. Through the synergistic effect of two factors, the exosome can reverse proliferation inhibition of DHT on hair follicle hair papilla cells, improve the cell migration ability, promote AGA mouse hair follicles to recover from a resting period to a growing period and block hair follicle microminiaturization, and finally reconstruct an AGA hair follicle microenvironment to achieve functional hair growth. Cell experiments confirm that the compound can repair hair follicle cells damaged by DHT, animal experiments show that the AGA mouse hair coverage rate is increased, the safety is good, the primary clinical effect is achieved, an efficient, safe and flexible new scheme is provided for AGA treatment, and the compound has good clinical transformation potential.

The application discloses a kind of immortalization human umbilical cord mesenchymal stem cell NOTCH1-LNR overexpression stable cell line construction method and its application in liver differentiation, it is related to immortalized cell technical field.The application first connects target gene to vector, and the slow virus plasmid containing target gene is constructed, the slow virus plasmid containing target gene is introduced into 293T cell, and high titer slow virus containing target gene is generated;Immortalized human umbilical cord mesenchymal stem cell is infected with slow virus, and mixed clone cell strain is screened, and the immortalized human umbilical cord mesenchymal stem cell NOTCH1-LNR overexpression stable cell line is obtained.The application proves that the differentiation efficiency of immortalized human umbilical cord mesenchymal stem cell NOTCH1-LNR overexpression can effectively improve liver-like cell to glycogen staining and Western blotting experiment.This research result is expected to provide new technical thought for the next step development to improve the yield of MSCs liver differentiation.

The present application relates to a porcine reproductive and respiratory syndrome virus infectious clone construction and rescue method, and belongs to the technical field of bioengineering. The porcine reproductive and respiratory syndrome virus infectious clone contains a full genome sequence of the porcine reproductive and respiratory syndrome virus, and the nucleotide sequence of the full genome sequence is the 26th to 15384th nucleotide of SEQ ID NO: 1 or a fragment having more than 80% identity with SEQ ID NO: 1 and having the same function. The present application also provides a plasmid pCAGGS-HA-CD163 expressing a porcine CD163 gene. By co-transfecting the aforementioned two plasmids into HEK-293T cells, the virus is rescued in vitro, a first porcine reproductive and respiratory syndrome virus infectious clone rescue platform based on HEK-293T cells is successfully built, and the porcine reproductive and respiratory syndrome virus is successfully rescued.

The invention discloses a method for drawing a human full-coverage protein interaction network based on digital PCR next-generation sequencing. After the advanced pedestrian 293T cell line obtains mRNA, a modified random primer or an oligo-dT primer is adopted to obtain a cDNA library; carrying out homogenization treatment on the human cDNA library; the method comprises the following steps: transforming plasmids of a BACTH bacteria double-hybrid system to obtain transformed plasmids; carrying out homologous recombination on the transformed plasmids of the sample library and the double-impurity system, and introducing into escherichia coli for screening to obtain positive PPI clones of the library with interaction; and carrying out digital PCR-based next-generation sequencing on the screened clones to draw the human full-coverage protein interaction network. The method is suitable for performing high-throughput screening after thousands of positive PPI combinatorial clones are obtained by'library-to-library 'bacteria or yeast double hybrids, identifying the same cell bar code cDNA combinatorial pairs, and realizing PPI network identification in different species, among species and in hybridization technology system extensive scenes.

The invention belongs to the field of gene engineering, and particularly relates to application of chikungunya virus structural protein in improvement of pseudovirus stability, a construction method of the chikungunya virus structural protein and vaccines, antibody evaluation and in-vivo gene therapy products. The method comprises the following steps: constructing a human codon optimized Asian strain CHIKV-E3 + E2 + 6K + E1 structural protein eukaryotic vector, and designing two types of binder expression plasmids of targeted T cells; the vector, a psPAX2 helper plasmid and a lentivirus target vector containing an SFFV promoter and a ZsGreen reporter gene (or a CAR therapeutic gene) are co-transformed into a 293T cell, and the lentivirus is prepared through culture, filtration and centrifugation. The lentivirus can efficiently infect Jurkat cells and activate human T cells, obviously reduces the infection efficiency on 293T cells so as to reduce the off-target effect, has excellent stability in human serum, and provides a safe and efficient gene delivery tool for in-vivo CAR-T therapy.

The invention discloses an FUT2 high-expression cell strain as well as application and a method of the FUT2 high-expression cell strain in vaccine quality control, and belongs to the technical field of vaccines. The technical problem to be solved is that a simple and easy-to-operate cell model is lacked in the prior art, and the cell model is used for evaluating the effectiveness of NoV vaccines on the cellular level. According to the key points of the technical scheme, the FUT2 high-expression cell strain is provided, the cell strain is an HEK 293T cell strain (HEK 293T-FUT2-EGFP) with EGFP fluorescence for overexpressing human alpha 1, 2-fucosyltransferase 2 (FUT2), and the cell strain stably expresses H, Lewis b and Lewis y type HBGA antigens; meanwhile, an alternative neutralization experiment of the cell level of the norovirus vaccine based on the cell strain is developed, a basis is provided for interaction research of the norovirus and host cells, and the cell strain has high practical application value.

The invention discloses a DNA (Deoxyribonucleic Acid) molecule for promoting mRNA (Messenger Ribonucleic Acid) expression and 3'UTR (Untranslated Region) in the mRNA molecule obtained by transcription of the DNA molecule, and the DNA molecule is derived from COX7B of a cytochrome C oxidase family. The invention also discloses an application of the composition containing the 3 'UTR in preparation of RNA (Ribonucleic Acid) therapeutic drugs or mRNA (Messenger Ribonucleic Acid) vaccines. Compared with a control group alpha-globin, the insertion of the 3'UTR fragment provided by the invention has the advantages that the expression quantity of renilla luciferase in Lent-X 293T cells is increased by 2.03 times, and the expression quantity of renilla luciferase in HeLa cells is increased by 3.62 times. The invention shows that the 3 'UTR has an excellent prospect of being applied to RNA (Ribonucleic Acid) therapeutic drugs or mRNA vaccines.

The invention discloses an AQP4-IgG (immunoglobulin G) detection method based on the combination of AQP4-M1 and M23 subtypes. According to the invention, two protein subtypes of AQP4-M1 and M23 are mixed in proportion by simulating physiological conditions and co-transfected to 293T cells, so that heterotetramers are formed and further assembled into a complete orthogonal array (OAP), and more antigen epitopes are exposed, thereby remarkably improving the recognition specificity and detection sensitivity of the AQP4-IgG antibody. Compared with traditional single subtype detection, the system can more truly simulate abundance and distribution of the AQP4 subtype in vivo, reduces false positive and false negative risks, and has a great value of rapidly and accurately screening AQP4-IgG related diseases clinically.

The present invention belongs to the technical fields of biomedicine and molecular biology, and particularly relates to a single-chain fragment variable (scFv) targeting human platelet-derived growth factor receptor (PDGFR)-β and use thereof in chimeric antigen receptor (CAR)-T cell immunotherapy. In the present invention, a scFv sequence targeting a human-derived PDGFRβ antigen is first obtained by immunizing a mouse, and then a second-generation CAR is constructed based on this, and additionally a CAR-T cell is obtained via lentivirus infection. The CAR-T cell can effectively kill a PDGFRβ antigen-positive 293T cell. The present invention provides a brand-new idea for eliminating PDGFRβ-positive cells to treat chronic kidney diseases, chronic liver diseases, cardiovascular diseases and various tumor diseases including various organ fibrosis, and has extremely attractive further development value and application prospects.

Provided in the present disclosure is a method for using a four-plasmid system to prepare modified immune effector cells. The method comprises: forming a lentivirus by using four plasmids within 293T cells, extracting and obtaining the lentivirus, then transfecting immune effector cells by using the lentivirus, and expressing a chimeric antigen receptor. Also provided in the present disclosure is a use of the immune effector cell obtained by using the described method and of a composition containing the immune effector cell.

The application discloses a method for constructing a cell membrane fusion model induced by a novel coronavirus-like particle, and belongs to the technical field of biology. The method comprises the following steps: constructing a virus-like particle containing a SARS-CoV-2 spike protein gene and an RNA packaging sequence PS9, using HEK-293T cells as packaging cells, co-expressing structural proteins M, E, N and S protein, and realizing assembly of VLPs without replication ability. Subsequently, the VLPs are used to infect immortalized type II alveolar epithelial cells, S protein expression is induced and cell-cell membrane fusion is mediated without the need of BSL-3 laboratory conditions. The formation of fused giant cells can be directly observed through ZO-1 protein immunofluorescence staining, the model is stable, has high biological safety, and is repeatable in vitro. The application overcomes the safety risk of live virus infection and the authenticity problem of a transient transfection model, can be used for research on a SARS-CoV-2 virus membrane fusion mechanism, and has a good application prospect.

This invention relates to a detection kit and preparation method for enhancing the detection efficacy of autoimmune encephalitis (CBA) antibodies. The kit comprises genetically engineered 293T cells stably integrating and co-expressing the eIF2α S52A mutant and HSP70, and transiently transfected with CBA-associated antigens. The preparation method includes constructing a co-expression vector, establishing a stable transfection cell line, stress preconditioning treatment, antigen transfection, and kit assembly. A method for assessing antibody affinity maturation using this kit is also provided. By synergistically modifying detection cells at three levels—translational regulation, protein folding, and anti-apoptosis—the invention significantly increases antigen expression, reduces apoptosis, and improves detection sensitivity and specificity. Furthermore, it integrates antibody affinity assessment into CBA detection for the first time, providing auxiliary information for clinical staging.

The invention provides a preparation method and application of fibronectin, and relates to the technical field of biology. The preparation method comprises the following steps: expressing fibronectin by using HEK-293T cells, and taking AGRN protein signal peptide as the signal peptide of the fibronectin. According to the preparation method, the HEK-293T cells can secrete and express fibronectin with high yield and high biological activity, and the application of the fibronectin in the aspects of tissue regeneration, wound healing and the like is facilitated.

The invention discloses a chimeric antigen receptor NK cell overexpressing a GPR133 gene, and a preparation method and application thereof, and belongs to the technical field of biological medicine. The preparation method comprises the following steps: (1) respectively mixing a pLL3.7-GPR133 plasmid with a packaging plasmid psPAX2, mixing a pLL3.7-NKG2D-CAR plasmid with a packaging plasmid psPAX2, mixing a pLL3.7-NKG2D-CAR plasmid with a packaging plasmid pMD2. G with a proper amount of an Opti-MEM culture medium; (2) mixing PEI with a proper amount of Opti-MEM culture medium, and incubating at room temperature; (3) adding the PEI-Opti-MEM solution in the step (2) into the Opti-MEM culture medium containing the plasmids in the step (1), and standing at room temperature; (4) adding the mixture obtained in the step (3) into 293T cells, uniformly mixing, and incubating at 37 DEG C; (5) collecting viruses from the cell supernatant, so as to respectively obtain a pLL3.7-NKG2D-CAR virus and a pLL3.7-GPR133 virus; and (6) co-transfecting an NK92 cell with the pLL3.7-NKG2D-CAR virus and the pLL3.7-GPR133 virus, so as to obtain the chimeric antigen receptor NK cell of the over-expressed GPR133 gene. According to the present invention, the GPR133 gene is subjected to overexpression in the CAR-NK cell for the first time, such that the biological function of the CAR-NK cell is significantly enhanced so as to improve the tumor cell killing ability of the CAR-NK cell.

The application belongs to the technical field of antibodies, and particularly relates to an antibody for specifically treating orf of milk goat kids and application thereof. Virus liquid of a diseased goat is collected from a lesion site, and ORFV virus strains are obtained through subculture and amplification of goat skin fibroblasts; milk goats are immunized with a weak vaccine to prepare high-titer immune serum; flow sorting technology is used to separate ORFV-specific B cells from PBMC (peripheral blood mononuclear cells); antibody genes are amplified through nested PCR, expression vectors are constructed, and 293T cells are transfected to express monoclonal antibodies. The monoclonal antibody No. 2 obtained in the application has high affinity and specificity, the heavy chain CDR1-3 region sequence is SEQ ID NO: 1-3, and the light chain CDR1-3 region sequence is SEQ ID NO: 4-6. The antibody can be used for diagnosis, treatment and prevention of orf of goats, and has the characteristics of high efficiency, specificity and stability, and provides a new biological preparation for prevention and control of orf of goats.

The invention relates to an mRNA nucleic acid molecule of an avian reticuloendotheliosis virus and application, and belongs to the technical field of immunology and biologica.According to the nucleic acid molecule, a T7 promoter, a 5 '-untranslated region (5' UTR), a gp90 antigen coding gene, a 3 '-untranslated region and poly (adenosine monophosphate) are used for constructing a recombinant plasmid. According to the invention, a gp90 gene sequence and polyadenylic acid are optimized to obtain the mRNA nucleic acid molecule of the reticuloendotheliosis of poultry, and the mRNA nucleic acid molecule is prepared into the mRNA vaccine; the constructed candidate mRNA vaccine is expressed in a 293T cell and is subjected to extracellular secretory expression.

The invention belongs to the technical field of targeted drugs, and particularly relates to a 293T exosome for targeted delivery of pirfenidone through a membrane-loaded FAP and a preparation method and application thereof.The exosome secreted by 293T cells serves as a natural nano-carrier, an FAP antibody with fibroblast targeting performance is modified on the exosome membrane surface through a copper-free click chemical method, and the exosome membrane surface is modified through a copper-free click chemical method; an anti-fibrosis drug PFD is efficiently loaded into the exosome by adopting an ultrasonic method, so that an engineered exosome system (293Texo antiFAP + PFD) with targeted recognition and drug delivery functions is constructed. The system is delivered to the lung in an aerosol inhalation mode, accurate enrichment and stable release of PFD at the pulmonary fibrosis focus can be achieved, and therefore lung tissue structure damage and collagen deposition are effectively relieved, and the anti-fibrosis curative effect is remarkably improved. According to the technology, the problems of low drug delivery efficiency, large side effect, poor targeting property and the like in traditional treatment are solved, and a safe, efficient and accurate treatment strategy is provided for pulmonary fibrosis.

The invention provides an EBV susceptible 293T cell capable of simultaneously expressing an epidermal cell receptor EphA2 and a cleavage replication key protein RTA, and an EBV susceptible BJAB cell capable of simultaneously expressing a B cell receptor CD21 and a cleavage replication key protein ZTA, so that the infectivity of the cells on EBV is remarkably improved. The EBV infection titer of the cell line disclosed by the invention is more than three times that of normal control under the same condition. The EBV susceptible cells are of great significance to research of EBV infection mechanisms, screening of antiviral drugs and research and development of vaccines.

The application discloses an IgA antibody targeting neurotrophic factors BDNF and NGF and application thereof. The IgA antibody targeting BDNF and NGF is obtained by expression and purification of expression plasmids thereof in HEK-293T cells, and the two IgA antibodies can specifically recognize and combine with BDNF and NGF respectively and have high affinity. The results of examples show that the IgA antibody targeting NGF and BDNF provided by the application can inhibit the growth, proliferation and survival of glioma cells, induce apoptosis of the glioma cells, and reduce the migration and invasion abilities of the cells, and the IgA antibody can also penetrate the blood-brain barrier and reach the glioma tissue to exert a significantly stronger anti-tumor effect compared with IgG antibodies.

The invention belongs to the technical field of biology, and particularly relates to a DNA aptamer for improving CRISPR / Cas9 mediated exogenous gene integration efficiency based on CtIP protein and application of the DNA aptamer, and the nucleotide sequence of the DNA aptamer is as shown in at least one of SEQ ID NO.1 and SEQ ID NO.10. The invention further discloses a preparation method of the DNA aptamer for improving CRISPR / Cas9 mediated exogenous gene integration efficiency based on CtIP protein. According to the invention, the key factor CtIP protein in DNA double-strand breakage repair is taken as a target for the first time, two aptamers are screened, and the donor vector of a CRISPR / Cas9 system is optimized by utilizing the aptamers, so that the integration efficiency of exogenous genes in HEK-293T cells and Hela cells is improved. The affinity of the CtIP protein specific DNA aptamer obtained through screening reaches the nanomole level, and the Kd value ranges from 42.56 nM to 407.29 nM. An aptamer is inserted into the tail end of a homologous arm of a donor carrier, and directional enrichment of a repair template is realized by utilizing an HMEJ mechanism. The aptamer A6913 and / or A13-32 significantly improves the integration efficiency of the exogenous gene, and the integration efficiency is improved by more than 3 times compared with a control group.

The invention provides a low-autophagy Vero cell line based on a DDIT3 inhibition gene as well as a construction method and application of the low-autophagy Vero cell line. The cell line stably knocks down DDIT3 expression through lentivirus-mediated shRNA, the autophagy level under the condition of nutrition deficiency is remarkably reduced, the cell proliferation capacity is enhanced, and the yield of influenza A H1N1 viruses is remarkably increased. The construction method comprises the following steps: cloning shRNA (short hairpin ribonucleic acid) to a GV493 lentiviral vector, co-transfecting 293T cell packaging virus with psPAX2 / pMD2. G helper plasmid, infecting Vero cells, and screening through puromycin to obtain a stable cell line. The cell line solves the problem of growth stagnation caused by over-activation of autophagy in serum-free suspension culture of traditional Vero cells, and is suitable for large-scale vaccine production.

This invention discloses a method for screening and validating personalized neoantigens for liver cancer and its application. DNA and RNA are extracted from tumor and adjacent normal tissue samples from patients and subjected to high-throughput sequencing. Mutant peptides are screened, and the patient's HLA typing is determined. The binding affinity of the mutant peptides to HLA alleles is then evaluated, and antigen presentation ability is predicted. Candidate neoantigen peptides are comprehensively screened. The candidate neoantigens undergo coding sequence sequencing, cloning, PCR amplification, in vitro transcription, capping and tailing modifications, and encapsulation to prepare an mRNA-LNP vaccine. The obtained mRNA-LNP vaccine is then transfected into 293T cells in vitro to verify antigen expression efficiency, completing the functional quality control of the mRNA-LNP vaccine. This invention provides a highly efficient, closed-loop system for screening personalized neoantigens for liver cancer and validating vaccines, an integrated approach that directly guides mRNA vaccine construction and in vivo immunogenicity evaluation from multi-omics data screening.

The invention belongs to the technical field of biology, and particularly relates to a universal type lentivirus titer detection method based on flow cytometry and application of the universal type lentivirus titer detection method. The detection method comprises the following steps: adding a transfection medium and lentivirus diluted by the transfection medium into 293T cells, and culturing; adding pancreatin for digestion to form a cell dispersion liquid; performing centrifugation and PBS resuspension on the cell dispersion liquid, adding a protein L antibody, performing incubation at 2-8 DEG C in a dark place for 30-60 minutes, then performing washing, and performing flow type detection on a CAR positive expression rate to obtain the lentivirus titer. The flow cytometry-based universal type lentivirus titer detection method provided by the invention is simple, convenient and rapid, has a relatively good linear range, and can accurately evaluate the titer of a lentivirus containing a CAR (Chimeric Antigen Receptor) gene.

The present invention belongs to the field of cellular immune engineering technology, and in particular to a kind of 293T multi-target target cell and its preparation method and application.The target cell includes a nucleotide fragment a expressing antigen CEA, a nucleotide fragment b expressing antigen CD19, a nucleotide fragment c expressing antigen BCMA, and a nucleotide fragment d expressing antigen CD70. Its preparation method is to select a basic cell (293T cell), construct it by means of exogenous construction, and then detect its antigen expression by flow cytometry, and sort out target cells that simultaneously highly express antigens CEA, CD19, BCMA and CD70 as target cells. The target cell exogenously expresses multiple antigens that can be used for the detection of multiple targets, and the operation that needs to be repeated 4 times is condensed into an operation once, which reduces the workload of cell culture, greatly saves time and cost, and the background factor secretion amount is normal during detection, the factor detection result is stable, the signal-to-noise ratio is higher, and it is more conducive to the quality detection of subsequent products.