89 results about "Bacterial outer membrane vesicles" patented technology

The present invention relates to the field of bioengineering, and in particular to a modified Bordetella pertussis strain, wherein the mltA gene and the pldA gene are mutated, thereby having an increased bacterial outer membrane vesicle (OMV) yield. The present invention also relates to a method for producing the modified Bordetella pertussis strain and a method for producing OMVs by the modified Bordetella pertussis strain.

The invention discloses a nano system for tumor radioimmunotherapy and a preparation method and application thereof, and belongs to the technical field of biological medicines.The system modifies bacteria outer membrane vesicles (OMVs) through engineering to enable the bacteria outer membrane vesicles to carry magnetite nanoparticles (Fe3O4NPs) and an ATR inhibitor VE822, the Fenton reaction activity of the Fe3O4NPs and the synergistic effect of the ATR inhibitor for inhibiting DNA damage repair are utilized, and the tumor radioimmunotherapy effect is improved. DNA damage is amplified, DNA damage reaction (DDR) is inhibited, generation of hydroxyl radicals (. OH) under radiation is enhanced, radiation resistance of tumor cells is reduced, immune reaction is reactivated, infiltration of effector T cells to tumors is improved, growth of mice tumors is remarkably inhibited, and an effective strategy is provided for tumor radioimmunotherapy.

The invention provides a nano photosensitizer modified by outer membrane vesicles of genetically engineered bacteria as well as a preparation method and application of the nano photosensitizer. The nano photosensitizer is a nano material with the particle size of 80-120 nm, and the nano photosensitizer modified by the genetically engineered bacterium outer membrane vesicle comprises dipalmitic acid phosphatidylcholine (DPPC), distearoyl phosphatidylcholine (DSPC), dioleoyl lecithin (DOPC), cholesterol, a small molecule dye and CD47nb-OMV membrane protein. The chemical structure of the small molecule dye is shown as a formula I or a formula II. The nano photosensitizer OBDlp modified by outer membrane vesicles of genetically engineered bacteria serves as a novel nano photosensitizer, tumor targeting PTT is mediated through NIR II FL / PA imaging, tumor immunogenicity is improved, antigen presenting cells are activated, and infiltration and functionalization of CD8 + T cells are caused. And OBDlp and laser are combined to induce release of a large amount of DAMPs and trigger ICD to promote maturation of cDC1s. Meanwhile, a CD47 nano antibody carried by the nano photosensitizer induces polarization of M1 phenotype macrophages by blocking a CD47-SIRP alpha signal transduction axis, and an immunosuppression tumor microenvironment is reversed.

The invention discloses a method for simultaneously extracting melanin and a bacterial extracellular vesicle compound from porphyromonas gingivalis and application of the method. The method comprises the following steps: inoculating a solid culture medium with a porphyromonas gingivalis seed solution, and carrying out anaerobic culture overnight at 37 DEG C; picking single colonies, putting the single colonies into a liquid culture medium, performing anaerobic culture at 37 DEG C for 24 hours, and then putting the single colonies into a bacterial incubator to be continuously cultured until OD600 is equal to 1; centrifugally separating supernatant liquid, and centrifuging again to obtain clarified supernatant liquid; and filtering the clarified supernatant liquid, carrying out ultrafiltration centrifugation on the obtained filtrate, washing a filter screen with water to obtain a washing liquid, carrying out ultrasonic treatment to obtain a melanin and bacterial exovesicle compound, carrying out vacuum freeze drying, and storing the melanin and bacterial exovesicle compound in a refrigerator. The method is economical, simple and convenient, the bacterial outer membrane vesicles and the melanin can be extracted at the same time, and the melanin and bacterial outer vesicle compound can achieve the effects of an immune agonist and a photo-thermal agent at the same time under 808 nm near-infrared laser irradiation.

The invention provides a bacterial outer membrane vesicle presenting Stoppin peptide as well as a construction method and application of the bacterial outer membrane vesicle, and relates to the technical field of biological medicines. According to the invention, a fusion protein ClyA-Stoppin is firstly designed and synthesized, and the fusion protein ClyA-Stoppin is displayed on the surface of the bacterial outer membrane vesicle by using a genetic engineering technology, so that the engineered bacterial outer membrane vesicle W-Stoppin OMV for targeting presentation of Stoppin peptide is successfully constructed. The W-Stoppin OMV provided by the invention has a remarkable inhibition effect on invasion and migration of tumor cells and can be used for remarkably promoting apoptosis of the tumor cells. According to the present invention, with the Stoppin monopeptide, the problems of insufficient Stoppin cell penetrability and difficult intracellular delivery are effectively solved, compared with the Stoppin monopeptide, the Stoppin monopeptide shows the excellent anti-tumor effect, and the migration inhibition, the invasion inhibition and the apoptosis promotion effects on the tumor cells are significantly increased. The W-Stoppin OMV provided by the invention has huge application potential in the aspect of preparation of anti-tumor drugs.

The invention discloses a bacterial outer membrane vesicle protein drug delivery system derived from attenuated salmonella as well as a preparation method and application of the bacterial outer membrane vesicle protein drug delivery system. According to the bacterial outer membrane vesicle protein drug delivery system, plug-and-play of a protein drug can be realized on the surface of a bacterial outer membrane vesicle. The invention constructs a bacterial outer membrane vesicle protein drug delivery system. OmpA of attenuated salmonella typhimurium VNP20009 is knocked out, recombinant OmpA of SpyTag inserted into an extracellular ring structural domain is supplemented, fusion protein of SpyCatcher and a protein drug is displayed on the surface of the bacterial outer membrane vesicle by means of the property that an isopeptide bond is spontaneously formed between SpyTag and SpyCatcher, the preparation method is simple and easy to operate, various proteins are displayed on the surface of the bacterial outer membrane vesicle, and the bacterial outer membrane vesicle has a good application prospect. And protein drugs are efficiently protected and loaded. And by combining the immunostimulation characteristic of the bacterial outer membrane vesicles, efficient treatment of tumors is realized.

The invention belongs to the technical field of drug co-delivery, and particularly relates to a nano-platform based on attenuated bacterial outer membrane vesicles and capable of achieving co-delivery of siRNA and adriamycin and a preparation method of the nano-platform. The preparation method comprises the following steps: synthesizing an ROS response type cationic polymer PTP; the preparation method comprises the following steps: preparing aptamer-DOX; preparing an ROS (reactive oxygen species) response type positive electric nano core PTP / 47aD; extracting and purifying the [delta] msbB OMVs; synthesizing a target functional unit DSPE-PEG2k-Angiopep-2 (Distearoyl Phosphate Polyethylene Glycol 2k-Angiopep-2); finally, the ROS response type co-delivery nano platform AO (at) PTP / 47aD is prepared and obtained. The co-delivery nano-platform prepared by the invention is relatively small in particle size, uniform in morphology and relatively high in biological safety, and has no obvious damage to various tissues and visceral organs. The glioblastoma immunopotentiator can be used as a general platform for gene delivery, can also be used as an immune activator, is suitable for intravenous injection administration, and can comprehensively overcome immune resistance of glioblastoma.

The invention belongs to the technical field of medicines, and particularly relates to production and application of a glucagon-like peptide-1 analogue-coated bacterial outer membrane vesicle which is mainly applied to treatment of type 2 diabetes mellitus (T2DM) and obesity. The glucagon-like peptide-1 (GLP1) analogue-coated bacterial outer membrane vesicle load gene has a coding sequence as shown in the GLP1 analogue of SEQ ID NO: 1. The bacterial outer membrane vesicles wrapping the GLP1 analogue, which are successfully prepared by the invention, have potential clinical applicability and research value in treatment of type 2 diabetes mellitus (T2DM) and obesity.

A plasmid capable of automatically inserting a membrane-localized luciferase reporter gene into a chromosome of a bacterium / recipient bacterium. After the plasmid inserts a luciferase reporter gene into a chromosome of a bacterium / recipient bacterium, the plasmid can be lost. Since the plasmid can be lost, the bacterium / recipient bacterium into whose chromosome the luciferase reporter gene is inserted does not require the addition of antibiotics during subsequent use, providing convenience and reducing interference.

The invention provides a bacterial outer membrane vesicle drug carrier with a targeting effect and preparation, and relates to the technical field of biological medicines.The bacterial outer membrane vesicle protein ClyA derived from an escherichia coli W3110 strain is combined with iRGD protein capable of being combined with an integrin receptor alpha v beta 3, a ClyA-iRGD-pGEX-6P-1 recombinant plasmid for expressing ClyA-iRGD fusion protein is constructed, and the ClyA-iRGD-pGEX-6P-1 recombinant plasmid is used for preparing the bacterial outer membrane vesicle drug carrier with the targeting effect. By endogenous expression of the plasmid in W3110 escherichia coli, the C-iRGD-OMVs which has a tumor targeting effect and shows ClyA-iRGD fusion protein on the surface is prepared. The C-iRGD-OMVs shows efficient targeting binding capacity to MCF-7 breast cancer cells, overcomes the defect that iRGD single peptide is prone to aggregation and crystallization, has the potential of loading drugs, lays a foundation for the C-iRGD-OMVs serving as a targeting carrier for delivering breast cancer treatment drugs, and is expected to be further developed into a targeting treatment preparation for breast cancer.

The invention relates to the technical field of biology, in particular to a preparation method and application of a bionic nano regulator. The invention provides a bionic nano regulator. The nano regulator comprises a therapeutic drug; the therapeutic drug is prepared from Ce6 and SB525334. And extruding the bacterial outer membrane vesicles and the PLGA encapsulating the Ce6 and SB525334 medicines to obtain the bionic nano regulator. Under photodynamic therapy, the bionic nano regulator can be recognized, taken and delivered to tumor tissues by neutrophils. The bionic nano regulator can regulate and control polarization of neutrophil to an anti-tumor N1 type, promote the neutrophil to release neutrophil elastase, specifically kill tumor cells and reverse a tumor immunosuppression microenvironment. Under the combined treatment of photodynamic-immunotherapy, the biomimetic nano regulator obviously inhibits tumor growth and enhances the anti-tumor immune response of the body.

The application provides a kind of bacterial outer membrane vesicle based on acid-sensitive peptide label and its preparation method and application, it is related to the technical field of engineered bacterial outer membrane vesicle.The ClyA-pHLIP fusion protein is designed and synthesized in the application, and the pHA-W3110OMV stably expressing the ClyA-pHLIP fusion protein is successfully obtained by using genetic engineering technology.The pHA-W3110OMV effectively solves the problem of easy aggregation of pHLIP in acidic environment, has good stability and solubility, and provides conditions for the stable targeting performance of pHLIP.Meanwhile, the pHA-W3110OMV not only retains the characteristics of targeting tumor cells in acidic environment, but also realizes the characteristics of targeting tumor cells in non-acidic environment.The pHA-W3110OMV has high targeting tracing efficiency on tumor cells under different pH conditions, and can realize qualitative and quantitative detection of tumor cells in peripheral blood, and has application potential in circulating tumor cell detection and tumor primary and metastasis localization.

The present invention is applicable to the field of biomedical technology, and provides a preparation method and application of multifunctional nanospheres targeting bacteria. The prepared targeted multifunctional nanospheres have both PDT / PTT functions and target harmful bacteria for antibacterial treatment of periodontitis. By using bismuth silver sulfide hollow nanospheres, photosensitizers and bacterial outer membrane vesicles as raw materials, the raw materials are cheap and the preparation process is simple. The prepared multifunctional nanospheres have unique advantages. They will not cause damage to probiotics, but can accurately target harmful bacteria and exert antibacterial effects, thereby effectively solving the problem of dysbacteriosis. In addition, the direct contact between the multifunctional nanospheres and bacteria will interfere with the electron transfer behavior of the bacteria, which is conducive to strengthening the bactericidal effect on bacteria. The adopted targeting strategy is extremely suitable for the treatment of dysbacteriosis diseases, showing a broad application prospect.

The present invention provides a method for preparing an antibiotic carrier based on bacterial outer membrane vesicles, characterized in that the preparation method comprises the following steps: (1) culturing bacteria in a culture medium containing antibiotics; (2) removing live bacteria from the culture medium and extracting the bacterial outer membrane vesicles, thereby obtaining an antibiotic carrier based on bacterial outer membrane vesicles; wherein the bacteria is a Salmonella mutant strain QS0074, whose deposit number is CCTCC M 2023192; and the antibiotic is acamycin. The antibiotic carrier based on bacterial outer membrane vesicles provided by the present invention has an excellent drug loading capacity for acamycin and good drug activity stability; the preparation method of the present invention is simple and easy to promote.

The invention provides an anti-tumor peptide modified bacterial outer membrane vesicle and a preparation method thereof, and relates to the technical field of engineering bacterial outer membrane vesicles. According to the invention, a recombinant plasmid capable of expressing a ClyA-A25 fusion protein in a prokaryote is designed and constructed, and the engineering bacterium outer membrane vesicle W-A25OMV modified by the anti-tumor peptide A25 is successfully prepared by using the recombinant plasmid. The W-A25OMV retains the basic structure of bacterial outer membrane vesicles, enhances the anti-tumor biological activity of A25 peptide, has good stability, shows a remarkable inhibition effect on invasion and migration of tumor cells, can effectively promote apoptosis of the tumor cells, and has development and application potential as an anti-tumor drug.

The invention discloses a bacterial outer membrane mixed vesicle carrier, the mixed vesicle is a mixed membrane structure formed by fusing two heterotype bacterial outer membrane vesicles, and the two heterotype bacteria are respectively gram-positive bacteria and gram-negative bacteria. The mixed vesicle carrier can simulate a natural internalization path of pathogenic bacteria (G < + > / G <->) invading cells, accurately locks cells which are easily invaded by bacteria, can identify corresponding homologous bacteria surfaces, efficiently enriches drugs at bacterial infection parts, and effectively improves treatment on mixed bacterial infection parts.

The invention discloses a fusion membrane packaged gold-iron core-shell nano diagnosis and treatment preparation as well as a preparation method and application thereof. The preparation takes gold-iron core-shell nanoparticles as a core, and the surface of the gold-iron core-shell nanoparticles is coated with a biomimetic membrane formed by fusion of bacterial outer membrane vesicles and tumor cell membranes. The preparation method comprises the steps of synthesis of the gold-iron nano core, surface functionalization and self-assembly coating of the fusion membrane. The preparation can simultaneously exert photothermal therapy and ferroptosis induction effects under near-infrared light irradiation, effectively initiates tumor cell immunogenicity death, and cooperatively activates dendritic cells and enhances T cell immune response by means of a pathogen related molecular mode carried by the fusion membrane and a tumor related antigen. Experiments show that the preparation can significantly inhibit tumor growth and metastasis, induce long-acting anti-tumor immune memory, and provide a multi-mode synergistic nano platform for tumor immunotherapy.

The invention provides a bacterial outer membrane vesicle based on genetic engineering modification as well as preparation and application thereof, and relates to the technical field of biological medicines. According to the invention, through a genetic engineering technology, a bacterial outer membrane protein ClyA from escherichia coli W3110 is combined with a tumor killing peptide KLAK, and a recombinant plasmid ClyA-KLAK-pGEX-6P-1 for expressing a ClyA-KLAK fusion protein is constructed; the plasmid is transformed into W3110 escherichia coli to realize endogenous expression of the fusion protein, and the engineered outer membrane vesicles C-KLAK-OMVs with anti-tumor activity are successfully prepared. On the basis that the original morphology of the bacterial outer membrane vesicle is reserved, the anti-tumor activity of KLAK is also reserved, migration and invasion of tumor cells HGC27 and MDA-MB-231 can be remarkably inhibited, and apoptosis of the tumor cells HGC27 and MDA-MB-231 is promoted; meanwhile, the medicine loading potential is realized; in addition, the C-KLAK-OMVs also improves the problem of insufficient stability of the KLAK peptide in serum, and provides a new thought and reference for targeted therapy of tumors.

The invention discloses an in-situ vaccine for tumor antigen capture driven by a nano motor as well as a construction method and application of the in-situ vaccine. The preparation method comprises the following steps: mixing prepared enzyme-immobilized dendritic silicon dioxide nanoparticles and drug-loaded bacterial outer membrane vesicles, extruding, carrying out solid-liquid separation, dispersing the obtained solid with a solvent, co-incubating the obtained suspension with pH-responsive membrane disrupting peptides and receptor molecules of targeted tumor cells, and carrying out freeze-drying to obtain the drug-loaded dendritic silicon dioxide nano-particles with the pH-responsive membrane disrupting peptides. The in-situ vaccine captured by the tumor antigen driven by the nano motor is obtained. The preparation method is green, simple and easy to operate. The in-situ vaccine can rapidly capture, enrich and release tumor-associated antigens and deliver the tumor-associated antigens to APCs, so that the reduction of curative effect caused by tumor heterogeneity and antigen degradation is avoided, the antigen presentation efficiency is remarkably improved, and the immune response is further enhanced; and the method is suitable for various tumors and has universality.

The invention discloses a nano-vesicle composition for inhibiting tumor metastasis after radiotherapy, which is characterized in that the nano-vesicle is an engineered bacterial outer membrane vesicle (OS-D), and the composition also comprises a pH responsive phospholipid modification component and a small interfering RNA (siRNA) capable of silencing CCDC25 gene expression; the siRNA is used for inhibiting formation of neutrophil Extracellular Traps (NETs) induced by radiotherapy and tumor cell migration and metastasis mediated by the NETs by blocking the interaction between the NETs and tumor cells, so that the effect of inhibiting tumor metastasis after radiotherapy is realized. The nano-vesicle composition provided by the invention can specifically block tumor metastasis mediated by neutrophil extracellular traps in a tumor microenvironment so as to enhance the effect of tumor postoperative radiotherapy and chemotherapy and effectively inhibit tumor metastasis and relapse.

The invention provides a preparation method of a drug carrier of paclitaxel nanoparticles wrapped in bacterial outer membrane vesicles, the preparation method comprising the following steps: (1) preparing outer membrane vesicles of Helicobacter pylori mutant strain QS003 (Helicobacteri pyloriΔfutBΔlpxEΔlpxFΔsncRNA‑2509025ΔtolB with plasmid Hbp+AffiHER2‑AffiEGFR1‑AffiVEGF); (2) mixing the obtained outer membrane vesicles with polylactic acid nanoparticles loaded with paclitaxel and mechanically extruding them to prepare outer membrane vesicles wrapped in paclitaxel nanoparticles. The drug carrier of paclitaxel nanoparticles wrapped in bacterial outer membrane vesicles obtained by the invention can well avoid the sudden release of the drug, the release ratio within 2 hours in the gastric fluid environment is less than 11%, the damage of gastric acid can be prevented, the degree of drug release in the stomach is reduced, and the drug utilization is improved.

The invention discloses separation and purification equipment for bacterial outer membrane vesicles, and belongs to the technical field of separation and purification equipment. The separation and purification equipment comprises a separation barrel and further comprises a first separation mechanism, the first separation mechanism is connected into the separation barrel and comprises a motor and a centrifugal barrel, and the motor is fixedly connected with the lower end of the separation barrel; the centrifugal barrel is fixedly connected with the output end of the motor and rotationally connected into the separating barrel, the upper end of the centrifugal barrel is fixedly connected and communicated with a feeding pipe, the feeding pipe rotationally extends out of the upper end of the separating barrel, a plurality of through grooves are formed in the periphery of the centrifugal barrel, first filtering membranes are fixedly connected into the through grooves, and a discharging hole is formed in the lower end of the separating barrel. The BEV separation device has the advantages that BEV separation can be achieved through the first filtering membrane in the first separation mechanism, preliminary separation and obtaining are achieved, materials are separated and purified through the second filtering membrane by means of the second separation mechanism, and the purification rate of the materials is effectively increased.

The invention discloses a composite microneedle with bacterial outer membrane vesicles loaded with active ingredients as well as a preparation method and application of the composite microneedle. The composite microneedle is prepared from the bacterial outer membrane vesicles and the active ingredients loaded in the bacterial outer membrane vesicles, the bacterial outer membrane vesicles are derived from mucus eubacterium, the active ingredient is cycloastragenol, and the composite microneedle has a transdermal delivery function and delivers the active ingredient to a target tissue in a targeted manner. The method comprises the following steps: firstly, encapsulating cycloastragenol into mucus eubacterium outer membrane vesicles through a co-incubation or active drug loading technology to form a composite nano-carrier; the EL-OMVs-CAG compound prepared by the invention has good biocompatibility and blood brain barrier penetrating ability, and can target a central nervous system, regulate the activity of microglial cells, inhibit neuroinflammatory response and reduce synaptic loss and neuronal apoptosis in a hippocampal region, so that cognitive function impairment caused by operation and anesthesia is improved.

The invention relates to a preparation method and application of an anti-Parkinson hydrogel preparation, solves the problems of oxidative stress of the brain of Parkinson and dysbacteriosis of intestinal flora, and adopts the technical scheme that the anti-Parkinson hydrogel preparation is prepared by the following steps: extracting bacterial outer membrane vesicles of alcaligenes faecalis through an ultracentrifugation method; the preparation method comprises the following steps: complexing cerium ions and polyphenol substances into a metal polyphenol network, loading the cerium metal polyphenol network into bacterial outer membrane vesicles by a mechanical extrusion method, and finally mixing the substances with inulin to obtain the anti-Parkinson hydrogel preparation. The preparation conditions are mild, the process is simple, the method is stable and reliable, the production cost is low, a new thought for treating the Parkinson's disease is developed, and the traditional Chinese medicine composition is an innovation for treating the Parkinson's disease and has huge economic and social benefits.

The application provides an E. coli outer membrane vesicle targeting TLR2 receptor and a construction method thereof, and relates to the technical field of biological medicine. The application first constructs a ClyA-HLYV fusion protein, and successfully obtains a recombinant plasmid ClyA-HLYV-pGEX-6P-1 stably expressing the fusion protein through genetic engineering technology, and induces endogenous expression of the fusion protein in an E. coli W3110 strain, to prepare a targeted bacterial outer membrane vesicle (HLYV-OMVs) expressing the ClyA-HLYV fusion protein on the surface. The HLYV-OMVs prepared by the application not only retain the specific recognition ability of HLYV to the surface TLR2 receptor of breast cancer cells, but also have the potential to encapsulate drugs, realizing the dual functions of targeted recognition and drug delivery. In addition, the HLYV-OMVs effectively solve the key problem of long-acting time of HLYV single peptide in targeted application, have good stability, significantly accelerate the effect starting speed after HLYV combines with the receptor, and lay an important foundation for efficient targeted treatment of tumors.

The invention provides a bacterial outer membrane vesicle capable of efficiently targeting tumors and a preparation method of the bacterial outer membrane vesicle, and relates to the technical field of biological medicines. The target protein PDL1 binding peptide is successfully modified on the surface of the bacterial outer membrane vesicles to prepare the bacterial outer membrane vesicles PDL1-OMVs, the particle size range of the bacterial outer membrane vesicles PDL1-OMVs is 20-250 nm, and the bacterial outer membrane vesicles PDL1-OMVs have an obvious saucer-like double-layer membrane structure and accord with the structural characteristics of the bacterial outer membrane vesicles. According to the PDL1-OMVs provided by the invention, on one hand, the defect that the PD-L1 binding peptide is weak in targeting efficacy is improved, the targeting capability of the PD-L1 binding peptide is remarkably improved, on the other hand, targeting function transformation is realized on the bacterial outer membrane vesicles, and the PDL1-OMVs can realize a specific targeting effect on tumor cells, so that conditions are provided for the PDL1-OMVs serving as a carrier for targeted delivery of tumor drugs.

The invention discloses a bacterial outer membrane vesicle. The bacterial outer membrane vesicle is obtained through induced expression and purification of a pseudomonas aeruginosa recombinant expression vector. The pseudomonas aeruginosa recombinant expression vector is obtained by knocking out an htrB1 gene on a wild type pseudomonas aeruginosa PAO1 strain; or the pseudomonas aeruginosa recombinant expression vector is obtained by knocking out the htrB1 gene and the tolB gene on the wild type pseudomonas aeruginosa PAO1 strain. The invention also discloses an application of the bacterial outer membrane vesicle in preparation of a pseudomonas aeruginosa vaccine. The pseudomonas aeruginosa vaccine can cause humoral immune response and cell-mediated immune response. According to the invention, a genome of pseudomonas aeruginosa is edited by utilizing a CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats-associated 9) technology, and an OMV attenuated pseudomonas aeruginosa strain and an OMV attenuated and productive pseudomonas aeruginosa strain are constructed. The OMV of the constructed strain is taken as an antigen, and broad-spectrum protective pseudomonas aeruginosa vaccines aiming at different serotypes can be developed.

The invention provides an escherichia coli outer membrane vesicle (OMVs-ClyA-NRP1) with a ClyA-NRP1 fusion protein displayed on the surface as well as a preparation method and application of the escherichia coli outer membrane vesicle, and relates to the technical field of biological medicines. The OMVs-ClyA-NRP1 effectively retains the tumor targeting activity of the NRP1 protein on the basis of maintaining the typical form and structure of the outer membrane vesicles of bacteria, shows excellent targeting binding capacity to breast cancer cells MDA-MB-231, and remarkably accelerates the interaction process of the NRP1 protein and tumor cells. Meanwhile, the vesicle system also has good drug loading potential. According to the invention, the bacterial outer membrane vesicles are modified through a genetic engineering technology, so that the bacterial outer membrane vesicles have targeted binding capacity with breast cancer cells, and a new strategy is provided for targeted therapy of breast cancer.

The present invention discloses a method for large-scale extraction of extracellular membrane vesicles (OMV / FGF2) secreted by Escherichia coli that has ampicillin resistance and can secrete bFGF protein after gene editing. The method cultured Escherichia coli in LB liquid containing 100ug / mL ampicillin, and used ultrasonic separation and filtration technology to effectively separate the extracellular membrane vesicles loaded with bFGF protein from Escherichia coli without Escherichia coli cells. The extraction method is simple and easy to operate, and the yield of extracellular membrane vesicles is greatly improved under the premise of retaining most of the effective biological components of bacterial outer membrane vesicles. At the same time, the method effectively removes the live bacterial cells of Escherichia coli, ensures the safety of the body, and greatly reduces the risk of bacteremia or scurvy induced by microorganisms entering the blood. In addition, the OMV / FGF2 extracted by the method was applied to colitis wounds, and it was found that it could significantly promote the healing of colitis wounds.

The invention belongs to the technical field of biological medicine and nano-drug delivery, and particularly relates to a long-acting growth hormone (hGH) wrapped bacterial outer membrane vesicle and application, and the outer membrane vesicle GH-OMVs wraps a human growth hormone mutant with an amino acid sequence shown as SEQ ID NO: 1. The GH-OMVs provided by the invention can be used for preparing medicines or preparations for preventing and / or treating diseases related to growth hormone deficiency or growth hormone-related metabolic disorder, including but not limited to growth hormone deficiency, dwarf / low height, osteoporosis and the like, and has a good clinical application prospect.