59 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 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.

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 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 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 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 provides a mesenchymal stem cell based on miR-148 as well as a preparation method and application of the mesenchymal stem cell. The preparation method comprises the following steps: inserting a miR-148 sequence into a lentiviral vector pLVX-Puro to construct a pLVX-miR-148 recombinant vector, packaging the pLVX-miR-148 recombinant vector in a 293T cell, and transferring the pLVX-miR-148 recombinant vector into a mesenchymal stem cell to obtain the mesenchymal stem cell based on miR-148. The mesenchymal stem cells based on miR-148 are umbilical cord mesenchymal stem cells with high expression of miR-148, the number of MSCs planted in the intestinal tract can be remarkably increased, the abundance of intestinal microorganisms and the number of lactobacillus reuteri are increased, and therefore the inflammatory bowel disease is improved.

The application discloses a method for drawing a human full-coverage protein interaction network based on digital PCR and second-generation sequencing. First, mRNA is obtained from a human 293T cell line, and then a cDNA library is obtained by using a modified random primer or an oligo-dT primer; then, the human cDNA library is uniformly processed; the plasmid of a BACTH bacterial double-hybrid system is modified to obtain a modified plasmid; the sample library and the modified plasmid of the double-hybrid system are homologously recombined and introduced into E. coli to screen positive PPI clones with interaction; and the screened clones are subjected to second-generation sequencing based on digital PCR to draw a human full-coverage protein interaction network. The application is suitable for high-throughput screening after obtaining tens of thousands of positive PPI combination clones by using a "library vs. library" bacterial or yeast double-hybrid system, and is suitable for identifying the same cell barcode cDNA combination to realize PPI network identification in different intra-species, inter-species and hybridization technology systems.

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 a COX17 gene 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 enables the expression quantity of renilla luciferase in LentiX 293T cells to be increased by 2.76 times and the expression quantity of renilla luciferase in HeLa cells to be increased by 6.41 times, and shows that the 3' UTR has an excellent prospect of being applied to RNA (Ribonucleic Acid) therapeutic drugs or mRNA (Messenger Ribonucleic Acid) vaccines.

The application belongs to the technical field of biological medicine, and provides a recombinant type III human collagen peptide for resisting HPV infection and a preparation method and application thereof, an amino acid sequence of which is a coding sequence of positions 991-1032 of type III human collagen UniProtKB: P02461 repeated three times, named as ATHPV3, and an amino acid sequence of which is shown as SEQ ID NO. 1. After expression and purification, the recombinant type III human collagen peptide with a molecular weight of about 13 kD is obtained, can be specifically combined with HPV 16 / 18 L1 protein, has multiple hydrogen bond actions, and can inhibit pseudo-virus infection of 293T cells at high and medium concentrations. The application successfully obtains a recombinant type III human collagen peptide with good purity, which can be combined with HPV 16 / 18 L1 and can effectively inhibit HPV 16 / 18 pseudo-virus infection of 293T cells, and provides an effective candidate drug for preventing and treating HPV infection.

This invention provides EBV-susceptible 293T cells that simultaneously express the epidermal cell receptor EphA2 and the key protein RTA for lysis and replication, and EBV-susceptible BJAB cells that simultaneously express the B cell receptor CD21 and the key protein ZTA for lysis and replication, significantly enhancing the infectivity of these cells for EBV. The EBV infection titer of the cell lines of this invention is more than three times that of the normal control under the same conditions. These EBV-susceptible cells are of great significance for studying the mechanism of EBV infection, screening antiviral drugs, and developing vaccines.

The invention discloses an MOG-IgG (Metal Organic Glycol-Immunoglobulin G) detection method based on a specific MOG subtype combined stably transfected cell. Two protein subtypes of MOG-alpha1 and MOG-beta1 are mixed and expressed in a 293T cell according to a ratio of 3: 1 to obtain a 293T-MOG-alpha1 + beta1 monoclonal stably transfected cell, and then a cytology-based determination (CBA) method is constructed on the basis of the monoclonal stably transfected cell and is used for detecting MOG-IgG. When the method is used for clinical serum MOG-IgG detection, the specificity is high, the sensitivity is high, the experimental result repeatability is good, the stably transfected cell strain can be stably subcultured, the consistency and the stability of the detection result are ensured, the detection efficiency is improved, and the occurrence probability of false positive and false negative is reduced.

The invention discloses an application of an exosome loaded with ENO1 protein in preparation of a product for treating female fertility decline. The method comprises the following steps: transfecting a lentivirus overexpression vector containing an ENO1 gene into a 293T cell, culturing, collecting a cell supernatant, and separating to obtain the exosome loaded with the ENO1 protein. Experimental results show that after the exosome delivers the protein ENO1, the number of primordial follicles, secondary follicles and sinus follicles can be remarkably increased, and the state of low ovarian function is improved; meanwhile, the mitochondrial function of granular cells is enhanced, and the energy metabolism level is promoted. A further research shows that the ENO1 can be specifically combined with mitochondrial outer membrane protein VDAC1, the mitochondrial membrane potential is stabilized, and the mitochondrial structure integrity of the oocyte is recovered, so that the energy metabolism in the oocyte is improved. Therefore, the invention provides a new treatment strategy for the fertility decline of the elderly women, and can be widely used for treating the fertility decline of the elderly women.

The invention discloses an application of an HEK-293T cell as a norovirus in-vitro culture host cell, the HEK-293T cell is used as a host bacterium for norovirus in-vitro culture for the first time, and meanwhile, intestinal bacteria such as enterobacter cloacae are introduced as an auxiliary factor, so that the norovirus in-vitro culture efficiency is effectively improved; and a stable and efficient human norovirus in-vitro culture system is successfully established. The defects of low norovirus culture efficiency and poor stability in the prior art are overcome, and stable replication and efficient harvesting of the norovirus are realized.

The invention discloses a bispecific antibody nanodisk as well as a preparation method and application thereof. The preparation method provided by the invention comprises the following steps: connecting a CD3 antibody gene and a PD-L1 antibody gene through a hydrophobic transmembrane region, cloning to a pCDH vector, packaging a lentivirus infected 293T cell, and screening through puromycin to obtain the 293T cell for stably expressing the CD3 * PD-L1 bispecific antibody; carrying out ultrasonication and gradient centrifugation to collect CD3 * PD-L1 bispecific antibody cell membrane vesicles; and mixing 22A polypeptide, the vesicles and DMPC according to a mass ratio of 1: 1: 5, rotating at a low speed of 3-5 DEG C, and uniformly mixing for 11-13 hours to obtain the bispecific antibody nanodisk. According to the scheme, the tumor targeting effect of the bispecific antibody can be improved, and the anti-tumor effect is enhanced.