147results about How to "High transfection efficiency" patented technology

The invention discloses an artificial antigen presenting cell for efficiently preparing a chimeric antigen receptor T cell (CAR-T) and a construction method of the artificial antigen presenting cell, and belongs to the field of cell therapy and immunotherapy, the artificial antigen presenting cell takes a K562 cell as a platform, an anti-CD3 single-chain antibody, an anti-CD28 single-chain antibody, a membrane binding type cell factor and a costimulatory ligand are stably expressed through genetic engineering, and the chimeric antigen receptor T cell (CAR-T) is obtained. And the local concentration of the anti-CD3 antibody is improved by adopting an HOTag3 tool. The multi-signal integration strategy obviously enhances the activation and proliferation capacity of the T cells, promotes the CAR-T cells to maintain a low differentiation state, and endows the CAR-T cells with excellent anti-tumor activity and durability. The artificial antigen presenting cell provides a novel efficient tool for the field of cellular immunotherapy, and is expected to promote development of more efficient and safer CAR-T therapy to cope with challenges of various malignant tumors.

The invention belongs to the field of biomedical materials, and particularly relates to a PBAE-DOPE-PEG2000 polymer-lipid nanocomposite and a preparation method and application thereof.The preparation method comprises the steps that a dimethyl sulfoxide-ethanol solution formed by mixing DOPE-PEG2000 lipid and PBAE serves as an oil phase, a sodium acetate / acetic acid buffer solution serves as a water phase, and the PBAE-DOPE-PEG2000 polymer-lipid nanocomposite is prepared through a microfluidic synthesis mode; the PBAE-DOPE-PEG2000 polymer-lipid nano composite material is prepared by the steps of carrying out ultrasonic treatment on the PBAE-DOPE-PEG The polymer-lipid nano composite material can efficiently load exogenous genes, the surface performance of the polymer-lipid nano composite material is changed and the stability of nano particles is improved by introducing DOPE-PEG2000 lipid, membrane fusion can be promoted, the risk of lysosome degradation is reduced, the transfection process of the exogenous genes in recipient cells is accelerated, and efficient expression in 293FT cells is realized. The efficient transfection of the polymer-lipid nanocomposite material is expected to show important application prospects in immune cell therapy or tumor cell therapy.

The invention relates to a preparation method of CAR-T cells and a product thereof. A method for preparing a CAR-T cell according to the present invention comprises the step of introducing a cyclic mRNA into a T cell that does not contain a chimeric antigen receptor; and, the cyclic mRNA comprises a first cyclic mRNA encoding a chimeric antigen receptor; in addition, the first annular mRNA comprises the following sequence (a) or (b): (a) a translation starting sequence and a coding sequence; and (b) translating the starting sequence, the coding sequence and the insertion element sequence. In the method, the transfection efficiency of the annular mRNA is high, and the positive rate is as high as 90%; compared with preparation methods of viruses, transposon systems and the like, the process is simple, the production cost is greatly reduced, and the safety is also improved; compared with linear mRNA, the protein expression time is greatly prolonged.

The invention provides nano-particles for generating CAR-T in vivo and application of the nano-particles, and belongs to the technical field of medical preparations. The nanoparticles are nanoparticles which are coupled with a CD7 antibody and entrapped with mRNA (messenger Ribonucleic Acid); the mRNA is obtained by transcription of a target gene; the target gene comprises a TROP2 single-chain antibody; the nucleotide sequence of the TROP2 single-chain antibody is as shown in SEQ ID NO: 1. The nucleotide sequence of the TROP2 single-chain antibody is optimized, the CAR containing the TROP2 single-chain antibody is constructed, then the CAR is transcribed to obtain mRNA, LNP entrapped with mRNA is prepared, then the CD7 antibody subjected to amino acid sequence optimization is adopted to be coupled with the LNP entrapped with mRNA, the transfection efficiency of the CD7 antibody in T cells is high, the CD7 antibody is injected into a mouse tumor model, and the lifetime of mice can be prolonged; t cells transfected by the T cell transfecting agent have high killing efficiency on HELA cells and LOVO cells.

The invention relates to a preparation method of a binary guanidyl block copolymer (Poly-PEG-GUA) for nucleic acid transfection of human vascular endothelial cells (HUVEC). According to the invention, N-(2-(3-carbamyl guanidyl) ethyl) methacrylamide with different proportions is subjected to reversible addition-fragmentation chain transfer (RAFT) polymerization to polyethylene glycol modified 4-cyano-4-(phenyl thioformyl sulfenyl) valeric acid (PEG-CPADN), so that five kinds of binary guanidyl block copolymers with different molecular weights are constructed; the HUVEC gene and messenger ribonucleotide (EGFP-mRNA) of green fluorescent protein are assembled in the HUVEC for screening of transfection efficiency, and the optimal molecular weight for HUVEC transfection is optimized. Experimental results show that the Poly-PEG-GUA with the molecular weight of 53575 Daltons can be used for effectively delivering green fluorescent protein (EGFP) mRNA (messenger ribonucleic acid) to HUVEC (human umbilical vein endothelial cells), and the highest transfection efficiency is shown. The optimized Poly-PEG-GUA in the invention has a certain reference value for the research and development of subsequent cardiovascular transfection products.

The invention discloses a photo-thermal cavitation micro-nano needle tip array, a manufacturing method thereof and a cell transfection method, and belongs to the field of cell transfection, the photo-thermal cavitation micro-nano needle tip array comprises a plurality of needle tip structures arranged on a substrate, the needle tip structures are located on the same side of the substrate, each needle tip structure comprises a main body, the main body is conical, an optical cladding and a photo-thermal layer are arranged on the outer surface of the main body, the photo-thermal layer is located at the top of the main body, the optical cladding is located between the photo-thermal layer and the main body, the main body is completely wrapped by the optical cladding and the photo-thermal layer, the photo-thermal layer is made of a photo-thermal material, the needle tip of the needle tip structure is in close contact with cells, and when photo-thermal cavitation bubbles are generated in the photo-thermal layer, the trepanning efficiency is higher, so that the cell transfection efficiency is improved; the positioning trepanning of cells is controlled through the arrangement and density of the needle tip array, and the functions of drug delivery, transfection, physical treatment and the like are achieved.

The invention relates to the field of nucleic acid editing, in particular to the technical field of regularly clustered interval short palindromic repeat (CRISPR). In particular, the present invention relates to Cas effector proteins, fusion proteins comprising such proteins, and nucleic acid molecules encoding them. The invention also relates to complexes and compositions for nucleic acid editing (e.g., gene or genome editing) comprising the proteins or fusion proteins of the invention, or nucleic acid molecules encoding them. The invention also relates to methods for nucleic acid editing (e.g., gene or genome editing) using the proteins or fusion proteins comprising the invention.

The invention relates to a TCR-T cell preparation method and a TCR-T cell product. The preparation method of the TCR-T cell provided by the invention comprises a step of introducing annular mRNA (messenger Ribonucleic Acid) into a T cell modified by a non-T cell receptor gene; and, the cyclic mRNA comprises a first cyclic mRNA encoding a T cell receptor; in addition, the first annular mRNA comprises the following sequence (a) or (b): (a) a translation starting sequence and a coding sequence; and (b) translating the starting sequence, the coding sequence and the insertion element sequence. In the method, the transfection efficiency of the annular mRNA is high; compared with a virus system and the like, the process is extremely simple, and the production cost and the detection cost are greatly reduced; compared with linear mRNA, the protein expression time is greatly prolonged; compared with a virus system, a transposon system and a CRISPR system, the safety risk is greatly reduced.

The invention discloses a yak intramuscular precursor fat immortalized cell line and a preparation method thereof. The method comprises the following steps: (1) obtaining yak primary intramuscular precursor fat cells; (2) performing SV40T lentivirus infection on the precursor fat cells in the yak muscle and screening cell strains: performing SV40T lentivirus infection by utilizing a 1 / 2 small-volume infection method to obtain immortalized cells after the precursor fat cells in the yak primary muscle are infected with SV40T, namely immortalized cells for short; puromycin is adopted to screen immortalized cells; and diluting the screened cells by using a limited dilution method, and carrying out enlarged culture to obtain the immortalized cell line. The yak intramuscular precursor fat immortalized cell line constructed by the invention has genetic homogeneity, the heterogeneity influence of cell populations is eliminated, and an in-vitro cell model is provided for subsequent research related to yak intramuscular fat cells.

The invention relates to a method for determining the biological activity of FGF21 (Fibroblast Growth Factor Receptor 21), which comprises the following steps: infecting and transfecting HEK293T cells by using an overexpression lentivirus containing FGFR1 and beta-Klotho genes and a reporter gene plasmid containing an SER reaction element so as to obtain a reporter gene-containing cell strain capable of stably expressing FGFR1, beta-Klotho and SRE; after FGF21 stimulation is added, the expression of a luciferase reporter gene at the downstream of the SER-containing promoter is activated; after co-incubation culture, adding a luciferase substrate, and fitting a four-parameter curve according to a measured reporter gene signal value, so as to determine the biological activity of the FGF21. According to the method provided by the invention, the test period is shortened to 24 hours, the detection sensitivity for FGF21 can reach a nanogram level and is improved by 1000 times compared with that of a traditional glucose oxidation method, and the result is consistent. The method provided by the invention can be used for evaluating the biological activity of FGF21 and analogues thereof as candidate drugs for treating metabolic diseases such as diabetes mellitus and NASH, and plays an important guiding role in quality control in research, development and production of FGF21 drugs.

The present invention relates to the technical field of nanopharmaceuticals, and in particular to a preparation method and application of fluorinated lipid nanoparticles. The preparation method of the fluorinated lipid nanoparticles comprises: dissolving fluorinated lipids and four commercial lipid molecules in anhydrous ethanol to obtain an ethanol phase lipid solution; dissolving nucleic acid in a citric acid buffer to obtain an aqueous phase nucleic acid solution; then uniformly mixing the ethanol phase lipid solution and the aqueous phase nucleic acid solution at a ratio of 1:3, and purifying the mixture through a desalting chromatography column to obtain the fluorinated lipid nanoparticles of the present invention. Compared with commercial transfection reagents and conventional lipid nanoparticles, the nanoparticles can significantly improve their nucleic acid delivery performance and transfection efficiency.

The present invention provides a polypolypeptide gene vector and a preparation method and application thereof, wherein the polypolypeptide gene vector comprises nanospheres formed by self-assembly of triblock polypolypeptides, wherein the triblock polypolypeptides comprise polyethylene glycol, polylysine and polyleucine connected in sequence; one end of the polylysine is connected to the polyethylene glycol via an amide bond, and the other end of the polylysine is connected to the polyleucine via a peptide bond. The polypolypeptide gene vector of the present invention can efficiently load the target gene in the polypolypeptide gene vector, and can form stable gene particles with high encapsulation rate and high load; the obtained polypolypeptide gene vector can produce a protonated sponge effect inside the lysosome to promote the lysosomal escape of the target gene and prevent the target gene from being degraded in the lysosome. It has important application prospects in the fields of gene editing, gene diagnosis, gene therapy and gene vaccines.

The invention relates to a method for efficiently establishing a stably transfected cell strain for knocking down and overexpressing a dyrk1b gene. Comprising the following steps: designing a specific RNA (Ribonucleic Acid) sequence to knock down a dyrk1b gene, constructing a DNA (Deoxyribose Nucleic Acid) sequence for highly expressing dyrk1b, and carrying out virus packaging and predetermined cell infection by utilizing a three-generation lentivirus system. A positive clone cell strain is obtained through Puromycin screening, OVCAR3 and 293T cells are infected by adopting a specific plasmid proportion, and knock-down and overexpression efficiency is verified. According to the method, the function of the dyrk1b gene is evaluated by observing the morphological change of OVCAR3 and COV362 cells, such as cell body elongation and antenna growth. The innovation point of the invention is that an efficient virus packaging system and precise gene expression regulation and control ensure the stability of the cell strain and the reliability of the experiment. In addition, the invention also relates to application of the stably transfected cell strains in comprehensive treatment of ovarian cancer magnetic hyperthermia, small molecule inhibitor and chemotherapy, and provides a new research tool and strategy for cancer treatment.

The invention relates to an exosome-like nano-vesicle for gene delivery and a preparation method thereof, the exosome-like nano-vesicle comprises a lipid phase and an inner water phase entrapped in the lipid phase, the lipid phase is prepared from cholesterol, phosphatidyl ethanolamine, phosphatidylserine, phosphatidylcholine and sphingomyelin; the inner water phase comprises nucleic acid drugs. The raw material components of the lipid phase are set, the use of cationic lipid and ionizable lipid is avoided, so that the anionic lipid vesicles simulating exosome lipid components can be prepared, the lipid phase entraps the internal water phase containing nucleic acid drugs, and the lipid vesicles have the characteristics of good stability, low toxicity and high transfection efficiency; the exosome has the advantages of limited immunogenicity, high delivery efficiency and enhancement of stability in the in-vivo circulation process of the natural exosome, the preparation process and the final quality are controllable, and commercialized production and clinical transformation are easier to realize.

The invention relates to the technical field of virus packaging. The invention provides a packaging method of a CAR (chimeric antigen receptor) virus. The packaging method comprises the following steps: mixing a plasmid pCMX-dR8.91, a plasmid pCMV-BaEV, a plasmid pAdVAnti and an opti-MEM culture medium, standing, mixing with a PEI transfection reagent, standing, mixing with a 293T cell, culturing, replacing the culture medium, continuously culturing, and harvesting the virus. A second-generation lentivirus packaging system is adopted, transfer plasmids are second-generation transfer plasmids needing to be activated by a Tat element, packaging plasmids are second-generation packaging plasmids pCMV-dR8.91 containing ag, pol, rev and tat, enveloping plasmids are second-generation enveloping plasmids pCMV-BaEV, plasmids pAdVAnti beneficial to improvement of transfection efficiency are added, and the packaging efficiency of the CAR virus is improved through the system.

The invention discloses a method for transfecting a plant receptor by using a cell penetrating peptide. The method comprises the following steps: (1) constructing an expression vector of a cell penetrating peptide fusion protein CPPs-Cys-mCherry; (2) transforming an expression vector of the cell penetrating peptide fusion protein CPPs-Cys-mCherry into a protein expression system; (3) inducing expression of cell penetrating peptide fusion protein CPPs-Cys-mCherry by using IPTG (isopropyl-beta-d-thiogalactoside) to obtain a crude protein sample; (4) purifying the crude protein sample obtained in the step (3), and then adjusting the concentration of the cell penetrating peptide fusion protein CPPs-Cys-mCherry to 1-200 [mu] g / ml; and (5) transfecting different types of plant receptors by using a solution containing the cell penetrating peptide fusion protein CPPs-Cys-mCherry obtained in the step (4), and completing the transfection.

The invention relates to the technical field of cell transfection experiment methods, and aims at effectively solving the cell liposome transfection tolerance phenomenon by increasing circulating cell hydrostatic pressure treatment in a cell liposome transfection experiment to improve the liposome transfection efficiency. In a cell liposome transfection experiment, the liposome transfection efficiency is improved by adding cell hydrostatic pressure treatment of a circulating mode; the circulation mode is that the hydrostatic pressure of the cells is maintained to be periodically changed in a set pressure interval value under a set frequency. The cell lipidosome transfection tolerance phenomenon is improved, and especially the transfection efficiency of lipidosome transfection tolerance cells such as primary culture cells and suspension culture cells is improved.

This invention relates to the field of gene vector technology, and more particularly to an ionizable comb-like polymer, its preparation method, and its applications. This comb-like polymer is a block polymer, comprising at least one block unit. Each block unit includes a first block, a second block, and a third block linked by covalent bonds. It exhibits high molecular flexibility and readily binds to nucleic acid-like substances to form polymer / nucleic acid complexes. Due to the presence of easily degradable intracellular glycosidic bonds in its structure, the polymer demonstrates high transfection efficiency. Furthermore, cell experiments show that the polymer / nucleic acid complex exhibits low toxicity to cells and significantly improves cell survival compared to the positive control. Therefore, it holds promise for widespread application in cell transfection. The preparation method provided by this invention utilizes a redox amination tandem reaction to directly prepare the target product, eliminating the need for intermediate product purification processes. This simple process facilitates large-scale production.

The invention discloses an anguilla japonica testis tissue cell AJTTC as well as an establishment method, a cryopreservation method, a resuscitation method and application of the anguilla japonica testis tissue cell AJTTC. The strain is preserved in the China Center for Type Culture Collection on January 22, 2024, and the preservation number is CCTCC NO: C202430. According to the method disclosed by the invention, the Japanese eel testis tissue is subjected to isolated culture by adopting a dry sticking method, then passage and cryopreservation are carried out, and the stable Japanese eel testis cell line is finally established, so that a new strategy mode and a research model are provided for researches on analysis of a Japanese eel germ cell differentiation mechanism, sex control, genetic breeding and the like.

The present invention relates to a PDRN derived from HEK293 cells and a preparation method thereof. The steps include: (1) centrifuging HEK293 cells, adding a lysate to obtain a lysate solution; (2) adding magnesium sulfate, protease and nuclease to the lysate solution, adding sodium dodecyl sulfate and sodium chloride after incubation, and obtaining a crude PDRN product after enzyme inactivation and alcohol precipitation; (3) dissolving the crude PDRN product in a sodium chloride solution, filtering, alcohol precipitation, washing, dehydration and drying to obtain PDRN. The PDRN preparation method provided by the present invention uses HEK293 cells as a source, uses a nuclease and a protease double enzyme one-pot method to prepare PDRN, has simple process control, does not have special environments such as high temperature, strong acid and alkali, and has relatively mild conditions. It does not need to add special equipment, and can be industrially produced on a large scale. In addition, the base composition ratio of the PDRN derived from HEK293 cells is completely consistent with that of the human body, has anti-inflammatory function and tissue repair activity, and has great application value in the fields of soft tissue repair, medical and aesthetic post-repair.

The invention discloses a preparation method and application of an iPS (induced pluripotent stem) exosome, and relates to the technical field of biologication.The preparation method comprises the steps of overexpression vector construction, nanoparticle preparation and transfection, iPS cell optimized culture, exosome separation, exosome purification, BMSC (bone mesenchymal stem cells) and exosome co-culture and bone defect repair experiment. The method has the advantages that the overexpression vector containing genes related to osteocyte differentiation, such as RUNX2 and ABC1, is constructed through gene editing, the vector is accurately introduced into the iPS cells by virtue of unique nano particle mediated transfection and magnetic field guidance, the transfection efficiency is greatly improved, the oxygen concentration is dynamically adjusted according to time in a culture link, and the osteocyte differentiation effect is improved. Meanwhile, substances such as a serum substitute and the like are added into the culture medium, a suitable growth environment is created for the iPS cells, the iPS cells are promoted to secrete high-quality exosomes, and the iPS cells are efficiently separated and purified by applying an advanced microfluidic technology and an affinity chromatography method, so that the yield and the quality of the exosomes are greatly improved.

The invention provides a separated Actin5C promoter as well as a construction method and an expression vector thereof, the separated Actin5C promoter is a partial region of a wild Actin5C promoter, and the partial region comprises a region from the 94th site in the downstream direction of a transcription start site to 400bp-600bp in the upstream direction. The separated Actin5C promoter is short in length, so that the expression vector containing the Actin5C promoter is small in size and high in transfection efficiency. By utilizing the expression vector provided by the invention, the target protein yield of in-vitro insect S2 cells can be remarkably improved, and the scientific research and clinical application values are high.

The application provides an isolated Actin5C promoter, a construction method and an expression vector thereof, wherein the isolated Actin5C promoter is a partial region of a wild-type Actin5C promoter, and the partial region comprises a region from the 94th site in the downstream direction to the 400bp-600bp in the upstream direction from a transcription initiation site. The length of the isolated Actin5C promoter is short, so that the size of the expression vector containing the same is small, and the transfection efficiency is high. The expression vector can significantly improve the target protein yield of insect S2 cells in vitro, and has high scientific research and clinical application values.

The invention discloses lentivirus transfection enhanced nanoparticles as well as a preparation method and application thereof, and belongs to the field of biomedical engineering. According to the invention, macromolecular polyethylene glycol with anions and a surfactant effect and micromolecular spermidine with cations and a cell phagocytosis enhancement inducing effect are used as raw materials for preparing the lentivirus transfection enhanced nanoparticles; nanoparticles with uniform particle size and high dispersity are prepared through electrostatic adsorption and low-temperature and low-speed self-assembly. After lentivirus is loaded on the nano-particles, lentivirus transfection enhanced nano-particles are obtained. The lentiviral transfection enhanced nanoparticles provided by the invention have very high transfection efficiency on tissue cells with relatively low cell membrane fluidity or relatively long growth cycle, have good biocompatibility, and can rapidly discharge cells after transfection, so that interference of empty carriers of the nanoparticles on subsequent research is reduced; therefore, the efficiency and accuracy of gene editing operation are greatly improved.

The invention discloses a liposome transfection method of chicken primordial gonad cells, and belongs to the field of cell transfection. The method comprises the following steps: rinsing chicken primordial gonad cells by using an OPTI-MEM culture solution, and then culturing the chicken primordial gonad cells; mixing and incubating the plasmid and the liposome transfection reagent to prepare a transfection mixture; wherein the plasmids comprise a donor plasmid and a PPARG (peroxisome proliferator activated receptor G) targeting plasmid; and mixing and incubating the transfection mixture and the primordial gonad cells. The gene editing positive cells are successfully obtained by optimizing the transfection process, and a large number of gene editing positive cells are obtained after amplification culture and flow sorting and can be used for preparing the gene editing chicken, so that the preparation time of the gene editing chicken is saved, and the preparation success rate of the gene editing chicken is increased.

The application discloses a cationic lipid molecule and a preparation method of the cationic lipid molecule for a lipid nanoparticle system, and provides a compound of formula I or a pharmaceutically acceptable salt thereof. In the application, a PEG lipid component is not contained in a three-component LNP composition, so that biological toxicity caused by the PEG lipid component is avoided, and safety is improved. Four-component LNP is simplified, and transfection efficiency is improved.

This invention belongs to the field of bioengineering technology, specifically relating to an improved transient transfection method for CHO-S cells. This transfection method includes the following steps: passage CHO-S cells for 3 days prior to transient transfection, adjusting the cell density to 0.35 × 10⁻⁶ cells / year. 6 Cells / mL were cultured in a shaker; on the day of transient transfection, the CHO-S cell density was adjusted to 4 × 10⁶ cells / mL. 6 -6×10 6 The cells / mL concentration was used to obtain a CHO-S cell suspension. Before transfection, the prepared solution A and solution B were mixed and allowed to stand to obtain a mixed droplet. The mixed droplet was added to the CHO-S cell suspension for culture to obtain recombinant CHO-S cells. The transfection method of the present invention not only has high transfection efficiency, but also obtains higher expression levels of the target antibody in CHO-S cells, providing a new approach for the development of protein biotherapeutic agents.

The invention relates to HIF-1alpha siRNA lipid nanoparticles and a preparation method thereof, and belongs to the technical field of pharmaceutical preparations. The preparation method comprises the following steps: S1, dissolving ionizable lipid, neutral lipid, cholesterol and pegylated lipid in an organic solvent to obtain an oil phase; the method comprises the following steps: dissolving HIF-1alpha siRNA in a first buffer solution to obtain a water phase; s2, mixing the oil phase and the water phase by adopting a micro-fluidic technology, diluting with a second buffer solution, removing the organic solvent, and concentrating to obtain the HIF-1alpha siRNA lipid nanoparticles, the targeted delivery of the HIF-1alpha siRNA is realized, so that the HIF-1alpha siRNA is effectively delivered to the rheumatoid arthritis diseased region through the delivery system to play a therapeutic role.

The invention provides a CRISPR-Cas9 near-infrared two-region precise light-operated editing system delivery carrier based on an erythrocyte membrane, a preparation method and application, and particularly, the CRISPR-Cas9 gene editing system delivery carrier provided by the invention has the advantages of low cytotoxicity, efficient transfection, high yield, targeting property, space-time controllability, easiness in large-scale production and multifunctionality. Compared with a traditional liposome transfection reagent, the carrier shows lower cytotoxicity, protects the CRISPR / Cas9 compound from being influenced by the in-vivo environment, and ensures that the CRISPR / Cas9 compound is smoothly delivered to target cells. By embedding a specific targeting receptor, efficient targeting delivery is realized, and the pertinence and effectiveness of treatment are improved. And by combining a near-infrared two-region material, the time-space precise control of a CRISPR / Cas9 system is realized, and deeper tissue penetrating power and smaller biotoxicity are achieved. The preparation process is simple, the method is suitable for industrial production and clinical transformation, and new possibility is provided for gene therapy and personalized medical treatment.

Provided is a method for modifying a cell, which comprises the following steps: transfecting a target molecule and a screening marker molecule to a cell in a non-viral manner, so that the cell expresses the target molecule and the screening marker molecule. Also provided is a modified cell obtained using the method. The method can effectively enrich the desired cell and increase cell positivity with respect to the target molecule.