30 results about "Membrane protein" patented technology

This invention provides a dengue / Zika virus genetically engineered vaccine and its application. The dengue / Zika virus vaccine comprises an open reading frame encoding envelope protein domain III (EDIII) and the non-structural protein NS1, and displays EDIII monomers in the delivery vector shell. Immunization with this vaccine primarily induces a type-specific antibody response, reducing the production of cross-antibodies and thus effectively avoiding or eliminating the risk of antibody-dependent enhancement of infection (ADE). This vaccine can be used to prevent dengue virus and Zika virus infection.

A targeting vehicles comprises an extracellular vesicle with a dopamine transporter antibody on a transmembrane protein of the extracellular vesicle, the extracellular vesicle is secreted by a cell transfected with a vector gene, and at least a portion of the vector gene comprises SEQ ID No: 1. The targeting vehicles provided in the present invention can be loaded with drugs and cross the blood-brain barrier to achieve specific binding to dopamine neuron, and regulate the secretion of Parkinson's disease marker proteins and delay the course of Parkinson's disease.

The present application relates to the field of biotechnology, in particular to a kit based on functionalized magnetic beads and application thereof in the field of stem cells. The functionalized magnetic beads of the present application are loaded with extracellular matrix proteins on the surface, and the extracellular matrix proteins are selected from collagen. The functionalized magnetic beads of the present application realize efficient capture and enrichment of collagen-bound transmembrane proteins, combined with mass spectrometry identification, and are suitable for screening stem cell transmembrane proteins, studying the action sites of stem cells and matrix, or developing drugs targeting stem cells. The kit and method of the present application have the technical advantages of simple operation, good stability and strong specificity, and provide a powerful tool for systematic study of the cell-matrix interaction mechanism mediated by stem cell transmembrane proteins.

The present disclosure is directed to compositions comprising mitochondrial optogenetics-based gene therapies and their methods of use. In some embodiments, a composition described herein comprises an expression vector comprising a first nucleic acid sequence encoding a channelrhodopsin fusion protein and a second nucleic acid sequence encoding a luciferase protein. In some cases, the first nucleic acid sequence and the second nucleic acid sequence are operably linked to an expression control sequence. In some instances, the channelrhodopsin fusion protein comprises a channelrhodopsin protein linked to an inner mitochondrial membrane-mitochondrial localization signal (IMM-MLS). In some implementations, when the expression vector is expressed, the luciferase protein is localized to the cytosol. In some cases, the IMM-MLS comprises a leading sequence from a mitochondrial inner membrane protein selected from ABCB10, ABCB140, Cytochrome C, and renal outer medullary potassium channel (ROMK).

Disclosed herein, in aspects, are compositions, methods, kits, and systems relating to delivery of payload into cells, for instance, for in vivo delivery via lipid delivery particles that comprise a chimeric envelope protein.

The application discloses a kind of bionic transmembrane protein affinity chromatography column and its preparation method and application.The system (iSTAC) realizes the in-situ construction of transmembrane protein in highly bionic dynamic microenvironment by integrating cell-free protein synthesis, functional mesoporous silica modification and amphiphilic (AH) peptide stabilized planar lipid bilayer technology.The core is to use long-chain PEG-24 crosslinking agent to retain lipid bilayer on the surface of silica gel, provide sufficient conformational dynamic space for multi-transmembrane protein, and introduce AH peptide to repair membrane defects, ensure that the receptor realizes directional embedding while maintaining natural functional attributes.The preparation cycle is shortened from 168 hours to 5 hours, which significantly improves the efficiency of targeted drug screening and in-situ analysis of binding sites.Using this platform, 5-HT 1A Receptor agonists crocin I and crocin II with anti-insomnia and neuroprotective effects are successfully screened from saffron, and the action site is accurately depicted.

The application belongs to the field of biology and relates to a nanobody targeting calexcin and a preparation method and application thereof, the nanobody targeting calexcin is successfully screened and prepared, and the nanobody-HRP fusion protein used for affinity identification in an enzyme-linked immunosorbent assay is successfully expressed; in combination with a phage-assisted continuous and discontinuous directed evolution (PACE) system, the nanobody screened is subjected to directed evolution, and stable mutation sites are found through sequence alignment analysis; it is proved through an indirect enzyme-linked immunosorbent assay and a luciferase complementation assay that the affinity of the nanobody after evolution to calexcin is improved by about 2 times.

The application discloses a kind of based on HIV envelope protein chimeric exosome and its preparation method and application, belong to biological medicine technical field.The application first constructs the cell line of stable expression HIV envelope protein Env, obtains engineered exosome from cell culture supernatant separation and purification;The exosome is used as immunogen combined with adjuvant immunization experimental animal, and high-efficiency induction specific humoral immune response;Again, obtain Env antigen specificity single B cell by flow cytometry sorting, obtain antibody variable region gene by single cell lysis, reverse transcription and nest PCR amplification, cloning to expression vector and expressing in mammalian cell, finally, HIV specific neutralizing antibody is screened by binding activity, affinity and neutralizing activity;The application is combined with single B cell antibody screening technology by embedding HIV Env antigen in the form of membrane combination in exosome surface, and realizes the synergistic optimization of antigen delivery and antibody screening process.

The present invention provides a central nervous system targeted pharmaceutical composition that can be used to treat various forms of brain damage, wherein an integrin antibody is attached to a transmembrane protein of an exosome through genetic engineering, and said exosome is used as a carrier to encapsulate a therapeutic drug.

This invention discloses a preparation process for a targeted drug delivery system based on the synergistic effect of Lamp2b and IMTP, comprising the following steps: constructing cells overexpressing the Lamp2b-IMTP fusion protein, culturing cells overexpressing the Lamp2b-IMTP fusion protein, thereby obtaining the targeted drug delivery system. The Lamp2b / IMTP synergistic targeted drug delivery system obtained by this invention allows for the specific accumulation of drugs in target cells, such as tumor cells, achieving precise drug delivery. The fundamental localization role of Lamp2b: Lamp2b is a naturally occurring transmembrane protein on the surface of exosomes, responsible for mediating the "initial recognition" between exosomes and recipient cells, ensuring that the delivery system, with exosomes as the core carrier, can be effectively recognized and bound by cells, avoiding rapid clearance by the body as a "foreign substance."

The present invention relates to a composition comprising a transfection agent and a human endogenous retrovirus (HERV) envelope protein having a mutation in its immunosuppressive domain (ISD) that reduces its immunosuppressive properties, or mRNA encoding at least the immunogenic portion thereof. The present invention also relates to the manufacture of pharmaceuticals and the use of this composition in therapeutic and prophylactic treatments.

The present application provides a central nervous system targeting pharmaceutical composition which can be used to treat various brain injury patterns, by genetically engineering an integrin antibody to bind to a transmembrane protein of an exosome, and coating the exosome as a carrier with a therapeutic drug.

Provided herein are compositions including a binding reagent for a Zika virus (ZIKV) epitope, pharmaceutical compositions including such binding reagents, and methods of detecting ZIKV binding antibodies and ZIKV neutralizing antibodies in patients.

The present invention relates to an IFITM scaffold enabling high-efficiency engineering of extracellular vesicles. In the present invention, active substances to be expressed in extracellular vesicles are linked to the IFITM scaffold so that the active substances can be expressed in the vesicles at a high level. In particular, the active substance linked to the C-terminus of the IFITM scaffold is present on the surface of the vesicles to exhibit the intrinsic activity. In addition, the present invention provides a U-shaped IFITM scaffold, which enables two types of active substances linked to the N-terminus and the C-terminus of the IFITM scaffold to be present on the surface of extracellular vesicles at a 1:1 ratio.

The present invention relates to the field of biomedicine. Disclosed are a target-switchable CAR-T cell linker system and a use thereof, specifically a target-switchable linker for improving the activation and proliferation of a specific T cell. The linker comprises a domain capable of binding to an antigen-specific T cell and a targeting domain capable of recognizing a cell surface membrane protein. On the one hand, the linker can link the antigen-specific T cell to APC to mediate the activation, phenotypic change, and proliferation of the antigen-specific T cell, enhance killing capacity, and reduce or reverse exhaustion; on the other hand, the linker can switch the CAR-T cell to a FRβ-targeting functional state, so that the T cell can recognize and kill an FRβ-expressing myeloid cell or macrophage, thereby achieving the activation, proliferation, and functional enhancement of the antigen-specific T cell. The present invention can significantly improve the sustained activation capacity and anti-tumor effect of CAR-T cells in a tumor microenvironment, reduce exhaustion, and enhance therapeutic safety.

The application discloses a kind of adipose tissue-derived mitochondrial-derived vesicles and its preparation method and application, belong to the field of biological medicine.The preparation method includes obtaining mouse adipose tissue, including at least one of interscapular brown adipose tissue, epididymal white adipose tissue, beige adipose tissue, in vitro cultured adipocyte;Mouse adipose tissue homogenate is removed by differential centrifugation Unbroken cell, nucleus, mitochondrial and organelle precipitate, the supernatant obtained is treated by trypsin and then ultracentrifuged, the precipitate is collected, washed, to obtain mitochondrial-derived vesicles, which are membrane-coated vesicles, with a particle size of 100-200 nm, enriched in mitochondrial membrane proteins and oxidative phosphorylation-related proteins, and proved by ischemic heart disease model that it can improve heart function, reduce the degree of ventricular remodeling in mice with ischemic heart disease and reduce the fibrosis area, for the preparation of drugs for preventing or treating ischemic heart disease, and has important clinical application value.

This invention discloses a sustained-release sulfated polysaccharide-based bioblended membrane, its preparation method, and its applications, belonging to the fields of biomedical materials and cosmetics technology. Addressing existing sulfated polysaccharide-protein blend systems, this invention utilizes silk peptides of specific molecular weights as bidirectional interfacial bridges, combined with a sulfated polysaccharide film-forming matrix, hydrolyzed eggshell membrane proteins, and moisturizing and plasticizing components to construct a quaternary synergistic system. This achieves stable and controllable construction of protein self-assembled topological phase-separated microdomains. These continuous network-like topological phase-separated microdomains can form an interpenetrating reinforced network and a molecularly compatible soft interface with the polysaccharide matrix, inhibiting harmful phase separation while comprehensively improving the membrane's mechanical properties, sustained-release controllability, bioactivity, and batch stability. All components are naturally biodegradable and free of chemical cross-linking, making it suitable for applications such as medical dressings and functional skincare patches.

The application discloses a chikungunya fever nucleic acid vaccine and belongs to the technical field of nucleic acid vaccines. A transmembrane protein sequence is obtained through multiple recombination optimizations, the protein sequence is connected with E2 and E1 protein sequences, a polyprotein is constructed, and the E2 and E1 proteins of chikungunya fever can be efficiently expressed through codon optimization and circular mRNA or linear mRNA technology. Compared with original virus protein sequences, the expression efficiency is improved by 2.7 times, the antibody titer of a mouse immunized by the circular mRNA vaccine is improved by 4.7 times, and the antibody duration is longer.

The utility model discloses an efficient enzymolysis reaction device for extracting pig intestine mucosa protein, which belongs to the technical field of animal product processing, and adopts the technical scheme that the efficient enzymolysis reaction device comprises a reaction tank, a heating mechanism is arranged on the inner wall of the reaction tank, a control mechanism is mounted on the left side of the heating mechanism, and a reaction cover is clamped at the top of the reaction tank; a servo motor is mounted at the top of the reaction cover, a connecting rotating rod is rotationally connected to the bottom of the reaction cover, a connecting rotating shaft is connected to the top of the connecting rotating rod through a flat key, stirring blades are welded to the surface of the connecting rotating rod, and a discharging pipe is fixedly connected to the bottom of the control mechanism. According to the heating mechanism, heat can be uniformly conducted into the reaction tank from the outer side of the reaction tank through cooperation of three heat conduction rings, a heat conduction rod and a heating ring, the local overheating or supercooling phenomenon caused by a traditional heating mode is avoided, the enzymolysis reaction is conducted in a stable temperature environment, the enzyme activity is guaranteed, and the extraction efficiency and quality of pig intestine mucosa protein are improved.

The present application relates to a kind of methods for identifying deep coverage based on near-infrared light catalysis mitochondrial proteome, in the presence of near-infrared fluorescent probe IR780 and capture reagent 3-ethynyl aniline (3-EA), mitochondrial protein is labeled in situ by using near-infrared light irradiation excitation, then, by click reaction, mitochondrial protein biotinylated labeled with IR780, and using streptavidin agarose microspheres, protein is enriched, and using liquid chromatography-mass spectrometry system, mitochondrial proteome is identified in depth coverage.The method uses near-infrared light catalysis adjacent label (PL-NIR), avoids the defect of visible light.Using the near-infrared excitation of IR780 and mitochondrial targeting, mitochondrial proteome is selectively labeled by spatially confined reaction in natural environment.PL-NIR strategy promotes the mapping of mitochondrial proteome, wherein up to 245 mitochondrial proteins are identified in live HeLa cells.PL-NIR strategy significantly improves the identification coverage, especially for the identification of mitochondrial inner membrane proteins.

The application provides a single cell analysis chip combining a carbon-based field effect transistor and microfluidics, comprising a plurality of single cell capture units; wherein the single cell capture unit comprises a sensing unit and a PDMS microfluidic channel unit arranged correspondingly to the sensing unit; the sensing unit is a carbon-based field effect transistor, and the PDMS microfluidic channel unit comprises a circular capture trap chamber vertically and directly above a sensing area of the carbon-based field effect transistor and a narrow channel; a first end of the circular capture trap chamber is connected with a first end of a first main channel, a second end of the circular capture trap chamber is connected with a first end of a second main channel through the narrow channel, and the first main channel and the second main channel are connected through an arc-shaped side channel; after cells are captured through the narrow channel, subsequent cells can flow into the next PDMS microfluidic channel unit through the arc-shaped side channel. The chip can separate and capture single cells, and achieve the purpose of detecting the surface proteins of single cells.

The application provides a double-layer guided bone regeneration membrane and a preparation method thereof, relates to the technical field of bone regeneration, and is composed of a compact layer and a porous layer which are arranged in a top-bottom stack, wherein the compact layer is prepared by cross-linking and modifying soluble eggshell membrane protein (SEP), epigallocatechin gallate (EGCG) and carboxymethyl chitosan (CMC) through genipin, has a compact non-porous structure and is used for blocking the migration of soft tissue cells; the porous layer is obtained by cross-linking hydroxyapatite modified by 3-aminopropyl triethoxysilane (APTES) and carboxymethyl chitosan (CMC) through genipin, has a three-dimensional porous structure and is used for promoting the growth of bone cells and bone regeneration. The double-layer guided bone regeneration membrane has good tensile strength, physical barrier function, antibacterial performance and controllable degradation, and can realize safe and effective bone formation effect under the irradiation of 808nm near-infrared light and 1W power.

The utility model discloses a blood sample collection card, including the card cover, collection paper strip, the card cover sets up at least one passageway for placing collection paper strip, and sets up the sample port on the passageway, collection paper strip includes blood filter membrane, PVC board and protein card, blood filter membrane, protein card set up on the PVC board, just the sample port is opposite blood filter membrane end when collection paper strip places the passageway, the utility model discloses a blood sample collection card made through collection paper strip inserts the card cover, effectively avoids the pollution of collection paper strip, realizes the storage to blood sample, and both carrying and using are convenient.

The present application provides a method for regulating the production of heterologous flavonoids and its application. The present application realizes the significant increase of flavonoids production by regulating the genes in the production cell, which includes up-regulating the transmembrane protein thiocyanate Ygap of E. coli or down-regulating one or more target genes. The present application also discloses the optimized host cell and its application.

This invention discloses a method for promoting the dissolution of eggshell membrane proteins and preparing chelated calcium through high-pressure crushing. The method includes the following steps: separating and drying the eggshell and eggshell membrane, then pulverizing them separately to obtain eggshell powder and eggshell membrane powder; adding water to the eggshell membrane powder and homogenizing under high pressure, followed by centrifugation to obtain an eggshell membrane solution; adding eggshell powder to the eggshell membrane solution, adjusting the pH to 1-3, and adding pepsin to react; after the reaction, adjusting the pH of the eggshell membrane solution to 6.5-7.5, and adding trypsin to react; after the reaction, inactivating the enzymes, centrifuging to collect the supernatant, adding anhydrous ethanol to the supernatant for alcohol precipitation, centrifuging to obtain the precipitate, which is chelated calcium. This invention overcomes the technical difficulty of dissolving eggshell membrane proteins, providing a new approach for the industrial extraction of soluble eggshell membrane proteins.

The application discloses a viral envelope protein, a lentivirus vector, a lentivirus and application thereof. The viral envelope protein is a mutant VSVG envelope protein, and the mutant VSVG envelope protein does not bind to a low-density lipoprotein receptor. The lentivirus vector comprises a nucleotide or a fragment thereof coding the envelope protein. The application specifically modifies the lentivirus envelope to adapt to the surface biological characteristics of NK cells and T cells, which is a core technical link for improving the transduction efficiency, reducing the activation threshold, and finally guaranteeing the safety and effectiveness of treatment. The application effectively enhances the efficiency and specificity of the lentivirus in NK cell and gamma-delta T cell infection by modifying the lentivirus envelope, has a wide application prospect in the field of adoptive immunotherapy, and provides practical basis and technical accumulation for developing the next generation of efficient and precise immune cell gene therapy tools.