173results about "Cell membrane modification" patented technology

The invention relates to the technical field of biological medicines, in particular to an iRGD targeted and anti-fibrosis small molecule dual-modified exosome as well as a construction method and application thereof. The exosome utilizes the specific binding penetration enhancement effect of iRGD (the amino acid sequence is CRGDKGPDC) and a fibroblast high-expression integrin receptor, so that the enrichment efficiency of the exosome at a focus part is improved; the anti-fibrosis micromolecule Let-7a-5p can inhibit a TGF-beta signal channel by blocking Smad2 / 3 phosphorylation, and the anti-fibrosis micromolecule Let-7a-5p and the anti-fibrosis microRNA carried by the exosome generate a synergistic anti-fibrosis effect; the exosome phospholipid bilayer can also protect Let-7a-5p from enzymolysis and prolong the half-life period, and iRGD modification can reduce the non-targeted uptake rate. An animal model verifies that the double-modified exosome has a good skin fibrosis resisting effect, and side effects such as liver and kidney function damage are not found. The invention provides an effective and safe treatment scheme for resisting skin fibrosis.

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

The invention discloses a prebiotic-loaded intestinal targeting exosome and a preparation method and application thereof.The preparation method comprises the steps that firstly, plasmids containing intestinal cell targeting peptide sequences are constructed through a gene editing technology, the plasmids are transfected and introduced into cells, the transfected cells are screened with antibiotics, and cell strains stably expressing intestinal cell targeting peptides are obtained; then culturing and adding prebiotics, and continuously culturing to enable the cells to take in the prebiotics and release the exosomes loaded with the prebiotics; and finally, collecting cell culture supernate, and centrifuging and resuspending for multiple times under a low-temperature condition to obtain the prebiotics-loaded intestinal targeting exosome. According to the preparation method, extraction of the exosome, loading of the prebiotics and an intestinal targeting modification process are optimized, so that the prepared intestinal targeting exosome loaded with the prebiotics can be used for remarkably improving the loading efficiency and intestinal targeting of the prebiotics and enhancing the stability of the exosome in gastrointestinal tracts; therefore, the delivery efficiency and the treatment effect of the prebiotics in the intestinal tract are improved.

The present invention provides compositions and methods for inducing antigen-specific tolerance in a subject. In one embodiment, the present invention provides a composition comprising an apoptotic body and an epitope of an antigen. Also provided herein are methods of preparing and administering the composition. The composition and methods provided herein can induce antigen-specific tolerance in a subject.

The present disclosure relates to a mammalian cell which is modified to express on the surface of its membrane a binding domain which binds to a target molecule. The disclosure also relates to protein constructs and nucleic acids for producing such modified mammalian cells, and to methods for using the mammalian cells to deliver therapeutic agents to target cells or tissues in vivo.

The present invention provides a method for safely and simply performing treatments such as exfoliation / floating treatment, dormancy treatment (dormancy not only enhances the protective effect but also suppresses unnecessary differentiation of cells, thereby maintaining an undifferentiated / low-differentiated state), and protective treatment for storage and transportation, on cells in tissue or cultured cells, while preventing their death and maintaining their viability, and a method for safely and simply performing cell activation treatment for implanting dormant floating cells. The cell treatment agent contains, as an active ingredient, at least one selected from alginic acid, heparins, sulfated dextran, and protease inhibitors.

Provided are: a culture medium which is for producing an organoid and which comprises a prostaglandin, hepatocyte growth factor (HGF), an interleukin (IL)-6 family protein, and an epidermal growth factor (EGF) family protein; a kit for cultivating an organoid; a method for producing an organoid; and an organoid.

The invention discloses an auxiliary protein composition, iron oxide nanoparticles, a preparation method and application, the composition comprises an SC protein with an amino acid sequence as shown in SEQ ID NO: 1 and an ST protein with an amino acid sequence as shown in SEQ ID NO: 2, and the composition can be used for modifying the iron oxide nanoparticles. The auxiliary protein composition can effectively promote endocytosis of the iron oxide nanoparticles, and assists the iron oxide nanoparticles to realize lysosome escape; moreover, the iron oxide nanoparticles modified by the auxiliary protein composition are good in biocompatibility and long in retention and storage time in cells, can be used for long-time labeling of the cells, and have wide clinical transformation and application prospects as a cell tracer agent.

The application provides a preparation method of organoids based on a single cell chip, which comprises microfluidic chip preparation, a two-water-phase solution preparation, a single cell suspension preparation, microfluidic manipulation, single cell-loaded microgel generation and organoid construction and the like. The application takes a microfluidic chip integrated with a normally closed pneumatic pump valve as a technical platform, takes a single cell suspension pre-incubated with a crosslinking agent as a chemical reaction core, takes a water-in-water droplet as a forming template, and realizes efficient in-situ loading of single dispersed seed cells in a micro hydrogel carrier through a one-step method, and simply removes empty droplets without cells in a subsequent transfer step. The single cells in the microgel can be cultured, expanded and self-organized to form required organoids. The application improves the consistency of the starting point of the organoids, has the high-throughput characteristics of the droplet microfluidic technology, realizes efficient and highly reproducible in-vitro construction of the organoids, and can play a great role in the fields of basic research and transformation application related to the organoids.

[Problem] To provide a method for producing a dendritic cell formulation that stimulates NKT cells, from monocytes collected from a recipient (monocytes collected by apheresis or peripheral blood containing monocytes), and that can exhibit the effect more quickly than conventional products. [Solution] This method for producing a dendritic cell formulation comprises: an adhesion step for placing collected monocytes in a culture vessel using a liquid culture medium, and leaving the culture vessel still to cause a portion of the monocytes to adhere to the inner surface of the vessel; a non-adhering cell removal step for removing non-adhering cells including monocytes other than the cells adhering to the inner surface of the culture vessel; a differentiation step for adding a predetermined factor in the culture vessel to cause the monocytes adhering to the inner surface of the culture vessel to differentiate into immature dendritic cells; a pulsing step for performing, with α-galactosylceramide, pulsing in the culture vessel in which the immature dendritic cells are present in a non-adhering state; and an NKT-stimulating dendritic cell induction step for inducing, from the immature dendritic cells, NKT-stimulating dendritic cells that stimulate NKT cells. The dendritic cell formulation is administered back into a recipient from which the blood have been collected.

Provided are a perfusion culture method and the like that are able to improve the productivity in the production of recombinant protein. Specifically, a method for improving recombinant protein production in perfusion culture of an animal cell having a capacity to produce a recombinant protein, the method comprising adding cystine and / or ammonium iron citrate to a cell culture medium, and the like, are provided.

There is provided a culture medium for expansion culture of a mammalian hematopoietic stem cell, where the culture medium contains a glutamate dehydrogenase inhibitor as an active ingredient.

The purpose of the present invention is to provide a novel coating film formation composition for the formation of a coating film having a function of inhibiting adhesion of a biological substance, a coating film using said composition, and manufacturing methods for a substrate for a cell culture, a multilayer film, and a cell aggregate using the coating film.　The present invention provides a coating film formation composition including: a polymer that includes structural units represented by formula (1) and formula (2) (in the formulas, R1-R3, X1, X2, T1, n1, n2, and n3 are as set forth in the specification and the claims) and that does not include a polysiloxane skeleton; and a solvent.

The present invention provides a method for culturing adherent cells, comprising a step for subjecting adherent cells to suspension culture in a medium including nanofibers composed of a water-insoluble polysaccharide, wherein the culture is conducted while stirring.

Compositions, systems, and kits are disclosed that comprise at least one immune checkpoint inhibitor and at least one immune cell type for adoptive cell therapy, wherein the at least one immune cell type has been fucosylated ex vivo (fuco-ACT). Also disclosed are methods of making and using the compositions, systems, and kits.

The invention relates to the technical field of biotechnology and immunotherapy, and discloses a method for inducing and amplifying pMHC specific homologous TSCM, which comprises the following steps: a) preparing pMHC presenting a single antigen peptide; b) sorting lymphocytes and mononuclear cells from a donor, and connecting the pMHC presenting the single antigen peptide obtained in the step a) to the surface of the separated mononuclear cells as stimulating cells; c) co-culturing the sorted lymphocytes serving as effector cells and stimulated cells in a culture medium containing a glycogen synthase kinase-3beta inhibitor, and inducing to generate pMHC specific homogeneous TSCM; and d) separating the pMHC specific homologous TSCM obtained in the step c). The method can induce and amplify sufficient pMHC specific homogeneous TSCM for adoptive immunotherapy, overcomes self tolerance and avoids or alleviates GVHD (Growth Vitamin Horse Disease); the preparation method is simple, induction and amplification efficiency is high, and universality and flexibility are achieved.

The present disclosure provides a method for chemoselective modification of a cell surface molecule on a target cell. A subject method includes contacting a target cell comprising cell surface molecule comprising a thiol, an amine, or an imidazole with a biomolecule comprising a reactive moiety, wherein the reactive moiety is generated by reaction of a biomolecule (e.g., an antibody) comprising a phenol moiety or a catechol with an enzyme capable of oxidizing the phenol or the catechol moiety. The contacting is carried out under conditions sufficient for conjugation of the cell surface molecule to the biomolecule, thereby producing a modified cell. The present disclosure provides kits for carrying out a subject method. The present disclosure also provides modified cells and methods for using same.

The invention belongs to the technical field of substance detection, and discloses a biomimetic nanomaterial coated with a cell membrane as well as a preparation method and application of the biomimetic nanomaterial. According to the invention, phospholipid functionalized magnetic nanoparticles are taken as a carrier, a cell membrane can be spontaneously integrated to the surface of the carrier by means of hydrophobic interaction, good binding capacity is achieved, and the obtained cell membrane-coated bionic nanomaterial is specifically alpha7nAChR-MNPs. According to the invention, a cell membrane for highly expressing an alpha7 nicotinic acetylcholine receptor is combined with phospholipid functionalized magnetic nanoparticles, so that alpha7nAChR-MNPs can specifically target potential risk substances, and the potential risk substances in a complex sample can be rapidly positioned and separated. The method is applied to food, potential risk substances can be predicted by simulating a biological recognition mechanism of a receptor, and the technical problems that a traditional screening method is insufficient in screening coverage degree, prone to causing misjudgment and missed judgment, high in cost and long in consumed time are solved.

The invention discloses an engineered exosome of LTH peptide modified miR-146a-5p and application, the core component of the medicine is the engineered exosome, and the engineered exosome is constructed by the following method: miR-146a-5p mimic infected human renal tubular epithelium HK2 cells with nucleotide sequences shown as SEQ ID NO.1-SEQ ID NO.2 are cultured and screened, the exosome rich in miR-146a-5p is separated from the culture supernatant of the cells, and the exosome rich in miR-146a-5p is obtained. Then, the kidney targeting peptide LTH is modified on the surface of the exosome. The invention reveals that HNRNP M protein is a key molecule for regulating and controlling miR-146a-5p to be sorted and enter the exosome for the first time, and verifies that the engineered exosome can be efficiently enriched in kidney, and by delivering miR-146a-5p to target IRAK1 gene in macrophage and inhibiting TRAF6 / IKK / NF-kappa B inflammation signal pathway, the macrophage is promoted to be polarized to a repairable M2 phenotype, so as to realize the purpose of improving the activity of the miR-146a-5p. And finally, the kidney injury induced by the calcium oxalate crystal is effectively relieved. The medicine has the advantages of clear mechanism, strong targeting property, good biocompatibility and the like, provides a brand new immunoregulation treatment strategy for renal injury, and has a wide clinical application prospect.

This method for delivering fine bubbles to cells is characterized by comprising a step for applying an electric field to a liquid containing fine bubbles, and thus delivering the fine bubbles to cells.

The present invention relates to processes and systems for preparing nanoparticles, cellular or viral membranes and / or cellular or viral membrane coated nanoparticles using or comprising, inter alia, a multi-inlet vortexing reactor, tangential flow filtration (TFF) and / or a high shear fluid processor such as a microfluidizer (or a microfluidizer processor). The present invention also relates to the nanoparticles, cellular or viral membranes and / or cellular or viral membrane coated nanoparticles prepared by the present processes and systems, and the uses and / or applications of the nanoparticles, cellular or viral membranes and / or cellular or viral membrane coated nanoparticles.

The present invention provides an oligodeoxynucleotide of unmethylated cytosine-phosphate-guanine dinucleotide and B lymphocytes bound thereto, which can target migration to diseased lymphoid organs and inhibit B lymphocytes activated under pathological conditions, thereby having a targeted therapeutic effect against pathological damage caused by B lymphocyte activation.

The present invention provides methods and means for producing nitrogenase polypeptides in the mitochondria of plant cells. The present invention provides methods and means for producing nitrogenase polypeptides in the mitochondria of plant cells.

The invention relates to a method for obtaining crystallised foetal bovine serum by adding agmatine that, at a specific concentration, acts to increase cell production and increase protein expression per cell. The method further relates to the specific form of reconstitution to liquid indicated in order to use same, without losing the original properties, allowing storage without an ultra-cold chain and reducing the risks of contamination, thereby allowing precise dosing of the amount to be used.

According to the targeting peptide and the engineered extracellular vesicle modified by the targeting peptide provided by the invention, the extracellular vesicle is modified by Pep-ACE2-2, so that the targeting property of the extracellular vesicle at an acute lung injury part is remarkably improved, and the acute lung injury is improved.

The present application relates to a kind of microcarrier and its application, the microcarrier has three-dimensional structure, the microcarrier has shell and inside is hollow or solid, the shell outer wall is uneven and has multiple concave points.This microcarrier improves the permeability of traditional microcarrier, is conducive to the material exchange between cell and external environment, improves cell survival rate, provides good mechanical protection for cell proliferation and constructs good three-dimensional structure for cell growth, realizes the friendly connection of cell, is conducive to the large-scale expansion of cell in limited space, and maintains the stability of cell physicochemical properties in long-term culture.

The invention discloses a modified exosome as well as a preparation method and application thereof, and relates to the technical field of biological medicines, CD44 targeted modification and composite active ingredients are synergistic to realize efficient repair of burn wounds and sensitive skin, and the modified exosome can be specifically enriched on the burn wounds or damaged sensitive skin parts through CD44 targeted peptide fragment coupling, so that the effect of repairing the burn wounds and the sensitive skin is achieved. The defects that a traditional repairing product is poor in targeting property and low in active ingredient utilization rate are overcome, the loaded dipotassium glycyrrhizinate, glutathione and FGF form a synergistic effect, wound healing and skin barrier repairing can be promoted through the FGF, the dipotassium glycyrrhizinate and the glutathione can be used for strongly resisting inflammation and oxidation, release of sensitive skin inflammatory factors is reduced, and the repairing effect is achieved. By matching with nitrogen protection incubation and a pulse ultrasonic loading process, the stability of active ingredients is greatly improved, finally, the repair effects that the healing process of burn wounds is remarkably accelerated, and sensitive skin percutaneous moisture loss is obviously reduced are achieved, and safety and effectiveness are both considered.

The present invention provides a herceptin antibody mutant. The herceptin antibody mutant is a VH-R50G, VH-R50A, VH-R50V, VH-R50L, VH-R50I, VL-F53L, VL-R66G, VL-H91Q, VH-R59T, VH-G103R, VH-F104L, VH-R50G / G103R, VH-G103R / F104L, VH-R50G / R59T / G103R, or VL-H91Q / VH-R50G mutant based on a wild-type light chain variable region (VL) and heavy chain variable region (VH). The mutant has lower antigen affinity, and ensures that effector cells can promptly dissociate from a target cell antigen upon exerting the cytotoxic effect thereof, thereby reducing self-apoptosis. Thus, the mutant exhibits enhanced tumor inhibition and / or killing activity and a prolonged in vivo half-life.

The invention discloses an engineered exosome for expressing a complete antibody and a preparation method of the engineered exosome, and belongs to the field of biotechnology and drug delivery. Aiming at the problem that a complete antibody is difficult to efficiently and stably express on the surface of an exosome membrane in the prior art, the engineering exosome with the complete antibody displayed on the surface is extracted from a culture supernatant after a target antibody and a natural antibody membrane-bound transmembrane region (TMD) sequence are fused, a recombinant lentiviral expression vector is constructed and a host cell is transfected. According to the method, TMD is used as an anchoring element, accurate positioning and efficient expression of an antibody on an exosome membrane are achieved, and the targeting recognition capacity of the exosome and the application potential of the exosome in the fields of targeting delivery, disease treatment and the like are remarkably improved. Experimental results show that the obtained exosome is typical in form, uniform in particle size and high in antibody expression efficiency, and a new technical scheme is provided for a targeting vector system based on the exosome.

Improved methods for large scale production of proteins and / or polypeptides in cell culture is provided. In accordance with the present invention, the method provides for culturing cells that have metabolically shifted. The use of such a method or system allows high levels of protein or polypeptide production and reduces accumulation of unwanted metabolic waste such as lactate. Proteins and polypeptides expressed in accordance with the present invention may be advantageously used in the preparation of pharmaceutical, immunogenic, or other commercial biologic compositions, such as antibodies.