35 results about "Transmembrane Region" patented technology

This invention provides a chimeric antigen receptor (CAR) targeting both BCMA and GPRC5D antigens and its uses. The chimeric antigen receptor (CAR) includes an extracellular localization signaling domain, an antigen domain targeting BCMA, an antigen domain targeting GPRC5D, a hinge region, a transmembrane region, a co-stimulatory factor, and an intracellular CD3ξ signaling domain. The antigen domain targeting BCMA includes a heavy chain variable region with an amino acid sequence as shown in SEQ ID NO: 1 and a light chain variable region with an amino acid sequence as shown in SEQ ID NO: 2. The antigen domain targeting GPRC5D includes a light chain variable region with an amino acid sequence as shown in SEQ ID NO: 3 and a heavy chain variable region with an amino acid sequence as shown in SEQ ID NO: 4. The dual chimeric antigen receptor, including an antigen domain targeting BCMA and an antigen domain targeting GPRC5D, can simultaneously recognize two anti-tumor targets, preventing tumor immune escape.

The present application relates to a kind of based on logic gate double-target point chimeric antigen receptor modified TIL cell, lentivirus expression vector and application, belong to tumor immunotherapy and gene editing technical field.The TIL cell based on logic gate double-target point chimeric antigen receptor modified in the application, double-target point chimeric antigen receptor includes chimeric antigen receptor EGFR and chimeric antigen receptor GD2;Chimeric antigen receptor EGFR is composed of CD8 alpha signal peptide, anti-EGFR single-chain antibody, CD8 alpha transmembrane region, 4-1BB costimulatory domain and CD3 zeta intracellular signal domain in series;Chimeric antigen receptor GD2 is composed of CD8 alpha signal peptide, anti-GD2 single-chain antibody, CD28 transmembrane region, CD27 costimulatory domain and CD3 zeta intracellular signal domain in series.The present application solves the defects that lentivirus transduction targeting is poor in prior art, CAR signal activation specificity is insufficient, TIL cell is easily exhausted, has the advantages that gene integration is accurate, signal transduction is controllable, in-vivo survival time is long, can be efficiently used for the immunotherapy of double-antigen co-expression solid tumor.

The object of this invention is to provide Labyrinthula microorganisms that have high sterol ester production capacity. [Solution] A Labyrinthull microorganism modified to have reduced or lost activity of sterol 24-C-methyltransferase (SMT1) compared to an unmodified strain, wherein the Labyrinthull microorganism is modified to express a modified diacylglycerol acyltransferase 2C (modified DGAT2C) gene, and the modified diacylglycerol acyltransferase 2C (modified DGAT2C) is modified to have a defect in presumed transmembrane regions 1 to 8 of the presumed transmembrane regions 1 to 12 in diacylglycerol acyltransferase 2C (DGAT2C).

Provided are a CD19-targeting humanized antibody and chimeric antigen receptor, and the use thereof. The humanized antibody contains CD19 VH and CD19 VL which are selected from one of groups 1) to 8). The CD19-targeting chimeric antigen receptor contains a CD19-targeting extracellular antigen recognition domain, a hinge region, a transmembrane region, and an intracellular domain, wherein the CD19-targeting extracellular antigen recognition domain contains CD19 VH and CD19 VL which are selected from one of groups 1) to 8).

This invention belongs to the field of protein engineering and biomedicine, and particularly relates to a water-soluble membrane protein, a recombinant vector, a recombinant host bacterium, and their modification methods and applications. The method involves the following steps: First, an interface mutant is constructed based on the SQTY code, and its water solubility and ligand binding ability are evaluated. If the requirements are not met, multiple low-impact transmembrane regions are screened, and after mutation modification, the interface mutant is introduced to construct a single-transmembrane combined mutant, whose water solubility and ligand binding ability are evaluated. If the requirements are still not met, the multiple low-impact transmembrane regions are combined in pairs, and the interface mutant is introduced to construct various double-transmembrane combined mutants, whose water solubility and ligand binding ability are evaluated, and the optimal double-transmembrane combined mutant is selected. This method rationally mutates CXCR4 in stages to achieve water solubility, minimizing changes to the protein's structure and other physicochemical properties, thereby maintaining or even enhancing its binding ability to the ligand CXCL12.

The application belongs to the technical field of biological medicine and molecular biology, and particularly relates to a CAR-macrophage with enhanced efferocytosis targeting apoptotic cells and a preparation method and application thereof. The CAR provided by the application comprises a signal peptide segment, a ligand recognition domain, a tag gene, a hinge region, a transmembrane region and an intracellular signal domain, can recognize lipid or protein signals exposed on the surface of apoptotic cells in an inflammatory microenvironment, and realizes targeted phagocytosis and aggregation in an inflammatory area. DKP type unsaturated ionizable lipids have protonation characteristics in an acidic microenvironment, which helps mRNA encapsulation and endosome escape. Lipid nanoparticles contain CAR mRNA and can respond to broken surface ligands. Under inflammatory conditions, the ligands fall off to expose DOPS, thereby improving the endocytosis capacity of macrophages. The CAR-macrophage and the nanoparticles can be used to prepare drugs for treating diseases such as metabolic-associated fatty liver disease and atherosclerosis, realize multiple synergies, have high specificity and safety, and have good industrialization prospects.

This application discloses a recombinant hemagglutinin protein, its uses, expression methods, and subunit vaccines, belonging to the field of biomedical technology. The technical solution is as follows: a recombinant hemagglutinin protein, obtained by removing the transmembrane and intracellular regions of the HA protein sequence of the H7N9-235 strain and fusing a T4-foldon sequence at the C-terminus; or by retaining the full-length sequence of the HA protein of the H7N9-235 strain. The amino acid sequence of the recombinant hemagglutinin protein is shown in SEQ ID NO.1 or SEQ ID NO.2. The application also discloses a recombinant hemagglutinin protein with good immunogenicity and medical prospects based on the H7N9 strain. Furthermore, when a subunit vaccine is prepared using the recombinant hemagglutinin protein with the transmembrane and intracellular regions removed, the immunogenicity after secondary immunization is significantly enhanced.

This invention provides a humanized antibody targeting CD19, a chimeric antigen receptor, and their uses. The humanized antibody comprises CD19VH and CD19VL, selected from groups 1) to 8). The chimeric antigen receptor targeting CD19 of this invention comprises an extracellular antigen recognition domain, a hinge region, a transmembrane region, and an intracellular domain targeting CD19. The extracellular antigen recognition domain targeting CD19 comprises CD19VH and CD19VL, selected from groups 1) to 8).

The present application relates to signal conversion receptors, and specifically provides a signal conversion receptor comprising an extracellular region, a transmembrane region and an intracellular region, wherein the intracellular region comprises a CD25 intracellular domain, and optionally further comprises an intracellular domain of a costimulatory signaling molecule. An immune effector cell expressing the signal conversion receptor has a higher positive rate, activation level and target cell killing ability compared with a control cell.

The application provides a virus envelope chimeric receptor and related biomaterials and applications thereof, and belongs to the technical field of molecular biology. Specifically disclosed is a virus envelope chimeric receptor, which comprises, in sequence, an antibody or antigen-binding fragment thereof, a hinge region, a transmembrane region and an intracellular region; the hinge region is selected from a hinge region composed of a hinge region of a CD8 molecule and a hinge region of a vesicular stomatitis virus envelope glycoprotein; the transmembrane region of the virus envelope chimeric receptor is selected from a transmembrane region of the vesicular stomatitis virus envelope glycoprotein; and the intracellular region of the virus envelope chimeric receptor is selected from an intracellular region of the vesicular stomatitis virus envelope glycoprotein. The virus envelope chimeric receptor is used for virus targeting, can improve the infection ability of viruses, reduces the infection rate of viruses on non-T cells to less than 5%, provides a safety guarantee for in-vivo CAR-T preparation, and can be used for industrial production.

This invention belongs to the field of biotechnology, specifically relating to a high-affinity anti-tumor NK cell, its preparation method, and its application. This invention designs NK92 cells transfected with a haPD1 high-affinity chimeric conversion receptor, wherein the high-affinity chimeric conversion receptor includes the extracellular segment of haPD-1, the transmembrane segment of CD28, the intracellular segment of DAP10, and the intracellular segment of CD3ζ. This invention prepares haChR3-NK92 cells through the construction of a recombinant lentiviral vector, lentiviral packaging, and lentiviral transfection of NK92 cells. In this invention, haPD-1 serves as the extracellular recognition domain of the CAR structure, specifically binding to PD-L1 on tumor cells to achieve a stronger tumor-killing effect. The co-stimulatory molecule CD28 serves as the transmembrane region to transmit extracellular information into the cell, integrating the adaptor protein DAP10 of the NK cell surface activator receptor NKG2D into the cell, and further embedding the intracellular segment CD3ζ commonly used in CAR design to jointly promote NK cell activation. The preparation method of this invention can successfully construct haChR3-NK92 cells, achieve the expression of the target plasmid, and be applied in anti-tumor drug research.

Provided are a cell membrane-expressed IL-15 fusion protein and a use thereof in cell therapy. The provided fusion protein comprises an IL-15Rα signal peptide, IL-15, an IL-15Rα sushi domain, and a transmembrane region; and a cell modified with the fusion protein can achieve efficient expression of IL-15 on the surface of a cell membrane, can effectively avoid side effects caused by the secretion and expression of IL-15, and is safe and controllable. An immune cell modified with a chimeric antigen receptor containing the membrane-expressed IL-15 fusion protein has a stronger tumor killing function, higher IFN-γ secretion, and a greater proliferative capacity; after treatment with lactic acid and TGF-β which are abundant in a tumor microenvironment, the immune cell retain a strong tumor killing function; and the immune cell can activate the anti-tumor function of immune cells other than the immune cell. The constructed immune cell can improve the therapeutic effect against tumors, prolong the duration of disease remission, and improve overall survival in patients.

This invention discloses a transgenic recombinant immune cell that specifically targets tumors. The recombinant immune cell comprises a chimeric polypeptide and a chimeric antigen receptor; the chimeric polypeptide includes a first extracellular region, a first transmembrane region, and a first intracellular region with HLA-G protein binding activity, wherein the N-terminus of the first transmembrane region is connected to the C-terminus of the first extracellular region, and the N-terminus of the first intracellular region is connected to the C-terminus of the first transmembrane region; the first transmembrane region includes the transmembrane region of the Notch receptor protein from Xenopus laevis; the antigen-chimeric receptor does not have HLA-G protein binding activity. This recombinant immune cell significantly improves the precise recognition of tumor cells by immune cells, reduces off-target effects of cell therapy, and accurately and effectively distinguishes tumor cells from normal cells, providing an effective target and targeting method for the treatment of broad-spectrum tumors (especially solid tumors).

This invention relates to the field of biomedical technology, specifically to a stabilized and modified herpes zoster virus gE protein, its preparation method, and its applications. The stabilized and modified herpes zoster virus gE protein of this invention is obtained through cysteine ​​mutation modification and structural design optimization, and exhibits improved expression levels, thermal stability, and immunogenicity compared to recombinant varicella-zoster virus gE protein with only the removal of the carboxyl-terminal hydrophobic transmembrane region and intracellular tail.

The application discloses a CD7 and CD3 double-targeting fusion protein and application thereof. Specifically, the application discloses an isolated fusion protein, which comprises, from N-terminal to C-terminal, (a) an anti-CD7 single-domain antibody (CD7 VHH); (b) an anti-CD3 single-domain antibody (CD3 VHH); and (c) a transmembrane region, which is used for anchoring the fusion protein on a cell membrane or a viral envelope. And a T cell-targeting pseudotyped lentiviral vector based on the fusion protein is constructed. The pseudotyped lentiviral vector of the application can effectively target and activate T cells, and can deliver the expression CAR-containing vector into T cells, thereby effectively activating T cells and enhancing the target killing ability of T cells.

This invention relates to the field of vaccine technology and provides a CHO cell line for exocrine expression of PEDV GIIb subtype S protein and its application. The CHO cell line integrates a codon-optimized PEDV GIIb S protein encoding gene carrying the TPA signal peptide into its chromosome. The nucleotide sequence of the PEDV GIIb S protein encoding gene is shown in SEQ ID NO.1. This invention uses currently prevalent GIIb strains as the antigen source to ensure vaccine strain matching; it constructs a secretory expression structure, thereby significantly improving the exocrine expression efficiency of the S protein; it precisely truncates the transmembrane region while retaining key structural domains, maintaining the protein's native conformation and immunogenicity; and it obtains a CHO cell line with stable gene integration and long-term passage to maintain expression levels through optimized transfection conditions and an antibiotic selection system.

The application provides a transfer plasmid of a lentivirus, a packaging system and application in preparation of the lentivirus, CAR-T cells and a kit, and particularly relates to a lentivirus packaging system for CAR-T, which comprises a transfer plasmid, the transfer plasmid comprises a virus promoter, a CAR promoter, a coding nucleotide sequence of a CAR expression region and a virus PolyA, wherein the virus promoter is selected from RSV, the CAR promoter is selected from SFFV and EF1 alpha promoter, the hinge and transmembrane region of the CAR expression region are from CD8a or CD8b, and the virus PolyA is selected from SV40 polyA. The lentivirus packaged by the packaging system has high packaging efficiency and transduction titer, so as to meet the demand of high production of the lentivirus for CAR-T cell treatment.

The application relates to the technical field of medical treatment and provides a CAR-gammadelta T cell targeting AXL and application thereof, the CAR-gammadelta T cell has a CAR molecule, the CAR molecule comprises the following structure: signal peptide-antiAXL scFv-extracellular hinge region-transmembrane region-co-stimulating factor-CD3 zeta intracellular region, the "-" is independently a connecting peptide or a peptide bond; the co-stimulating factor comprises a CD28 intracellular region and a 41BB intracellular region. Advantages: the CAR-gammadelta T cell targeting AXL integrates a third-generation CAR structure (CD28+4-1BB double co-stimulating domain) and a gammadelta T cell, realizes high targeting precision and killing efficiency. The in-vitro killing rate of the CAR-gammadelta T cell targeting AXL to AXL positive lung cancer cells (A549 and HCC827-ER3) is >90% (effector to target ratio 10:1), is obviously improved compared with common gammadelta T cells, and the tumor volume is obviously reduced in a PDX mouse model, which proves excellent treatment effect.

A dual-targeting chimeric antigen receptor (CAR) and its encoding gene, a recombinant expression vector, natural killer cells, and applications thereof are provided, relating to the field of tumor immunotherapy. The dual-targeting CAR includes a dual-CAR molecular encoding sequence sequentially connected encoding sequences of: an extracellular antigen-binding region, a transmembrane region, and an intracellular signal transduction region. PD1 molecule extracellular domain and NKG2D molecule extracellular domain serve as a first recognition target and a second recognition target respectively. When the PD1 molecule extracellular domain recognizes PD-L1 molecules on tumor cells, the negative regulatory signal is transmitted downward, triggering activation of the DAP10 activating receptor pathway and converting it into a positive activation signal. Simultaneously, when the NKG2D molecule extracellular domain recognizes NKG2DL molecules on tumor cells, NK cell activation is induced. Under the synergistic action of costimulatory molecules 4-1BB and CD3ζ, the anti-tumor therapeutic efficacy of NK cells is significantly enhanced.

The present application relates to the technical field of biological medicine, in particular to a kind of stable modified varicella-zoster virus gE protein, its preparation method and application.The stable modified varicella-zoster virus gE protein of the present application is obtained by cysteine mutation modification and structure design optimization, compared with the recombinant varicella-zoster virus gE protein with only carboxy-terminal hydrophobic transmembrane region and intracellular tail removed, has improved expression, thermal stability and immunogenicity.

The present application relates to a kind of encoding aspergillus fumigatus chimeric antigen circular RNA and its application, belong to biomedical and vaccine technology field.The present application first applies circular RNA technology to anti-aspergillus fumigatus infection, provides new strategy for fungal vaccine development.Select the non-transmembrane region of aspergillus fumigatus cell membrane protein FtrA to construct antigen, avoid the transmembrane region expression problem and immune recognition uncertainty;Fusion IgK signal peptide and Fc domain, significantly improve antigen secretion efficiency, stability and immune presentation effect.LNP delivery system mature, efficient, can effectively deliver vaccine to host cell and express antigen, the LNP@CircRNA provided by the present application, especially the LNP@CircRNA FtrA2 , can induce high level of IgG antibody and T cell immune response in mouse in vivo without using adjuvant, realize the dual activation of humoral immunity and cellular immunity.

The application discloses a preparation method of rabies virus glycoprotein and application thereof, and belongs to the technical field of biological medicines, and comprises the following steps: expressing a glycoprotein G gene of rabies virus containing an extramembrane region, a transmembrane region and an intramembrane region in yeast; performing cell disruption on the yeast, adding a detergent, and obtaining a solution containing rabies virus glycoprotein G; purifying the solution to obtain full-length rabies virus glycoprotein G; and immunizing the prepared full-length rabies virus glycoprotein G through intramuscular injection, oral administration or atomization inhalation. The scheme provided by the application has the advantages of strong immunogenicity, high safety, low production cost and easiness in scaling, solves the problems of complex traditional vaccine process and high cost, and provides a reliable new type of subunit vaccine solution for rabies prevention.

The present application provides a kind of CD47 mutant and its application, wherein CD47 mutant includes the amino acid sequence of partial amino acid sequence of CD47 and glycosyl phosphatidyl inositol (GPI) connection signal.The CD47 mutant is obtained by removing potential cell survival inhibition, blood flow and angiogenesis inhibition and other negative effect signal transduction sequences (transmembrane region and intracellular region) in CD47, while the beneficial functional segment (extracellular IgV domain) involved in inhibiting the phagocytosis of myeloid cells and its cytotoxic activity in CD47 sequence is fused with GPI membrane anchor connection signal sequence from human decay accelerating factor (DAF) protein.The obtained CD47 mutant can protect cells and organ transplants from being mediated by myeloid cells and other SIRP alpha positive immune cells with equal efficacy to wild type CD47, and at the same time eliminates the negative effects of wild type CD47.

The present disclosure provides bispecific chimeric antigen receptors targeting CD20 and BCMA. The CAR may comprise an scFv targeting CD20 and an scFv targeting BCMA, a hinge region, a transmembrane domain, a co-stimulatory region, and a cytoplasmic signaling domain. The chimeric antigen receptors can be used to treat autoimmune disorders or cancer.

The application discloses a kind of chimeric antigen receptor macrophages and preparation method thereof.To the problem that traditional T cell costimulatory domain is not matched with macrophage signal pathway, the application provides a CAR structure specially designed for the biological characteristics of macrophage.The chimeric antigen receptor expressed by the CAR-M cell includes at least one costimulatory domain selected from Ncr1, TLR5, Rage and Tim4, and the CAR structure is composed of CD22 scFv extracellular antigen recognition region, CD8 alpha hinge region, transmembrane region, the costimulatory domain and CD3 zeta intracellular signal domain in sequence.By constructing the recombinant expression plasmid of pB vector skeleton and inserting P2A-EGFP expression monitoring element, RAW264.7 macrophages are transfected by using Zeta life transfection reagent, and the CAR positive rate can reach more than 97% by flow cytometry or confocal microscopy verification.The application realizes the efficient activation of phagocytosis, antigen presentation and microenvironment remodeling functions specific to macrophages, and significantly improves the application potential of CAR-M in solid tumor treatment.

This invention discloses a GPC3-targeting CAR-NK cell, its preparation method, and its applications, belonging to the fields of immunology and molecular biology. The CAR-NK cell stably expresses a chimeric antigen receptor (CAR). The CAR structure, from N-terminus to C-terminus, consists of a signal peptide, a GPC3-targeting single-chain antibody scFv, a CD8a hinge region, a CD8a transmembrane region, a 4-1BB intracellular region, a modified CD3ζ intracellular region, and human IL-15 linked via a P2A peptide. The GPC3-targeting single-chain antibody scFv is formed by linking the light chain variable region (VL) and heavy chain variable region (VH) of a GC33 single-chain antibody via a G4S linker, with the linking order being VL-VH. The modified CD3ζ intracellular region incorporates a YXXQ structural motif only at the site closest to its C-terminus. This invention can completely eliminate tumors in a mouse orthotopic hepatocellular carcinoma model without recurrence, exhibiting high efficacy, high safety, and high accessibility, and can be used to prepare drugs for treating hepatocellular carcinoma.

The present application relates to a kind of dsRNA of prawn SGLT1 gene and its application.The target sequence of the dsRNA is derived from the coding sequence corresponding to the transmembrane region outside loop, intracellular loop or extracellular loop of sodium-dependent glucose cotransporter SGLT1 gene coding protein.The dsRNA of prawn SGLT1 gene provides a new molecular means for preventing and controlling acute hepatopancreas necrosis disease.The dsRNA molecule can be synthesized by in vitro transcription or prokaryotic / eukaryotic expression system, and can be further constructed to contain the sequence expression cassette, recombinant vector and recombinant bacteria.The dsRNA molecule of the present application is small in molecular weight, biodegradable, and will not cause environmental pollution or drug residue, with the advantages of safety, environmental protection, high efficiency and scale production, etc., and can be used as a new biological agent or feed additive for preventing and controlling Vibrio parahaemolyticus AHPND infection.

The application belongs to the technical field of nanometer hole single molecule detection and biochemistry, and provides a nanometer hole based on zebra fish protein and application thereof in single molecule detection. The zebra fish protein is wild type JAC5 protein, and the nanometer hole based on the wild type JAC5 protein is applied to single molecule detection. Compared with other nanometer hole proteins, the wild type JAC5 protein has significant advantages in cloning expression and purification process, and is suitable for detecting nucleic acid, protein and pathogen, and has wide application scenarios. Compared with the wild type JAC5 protein, the mutant zebra fish protein after amino acid mutation of the wild type JAC5 protein can normally express, enhance structural stability and improve detection performance by eliminating the steric hindrance of the sugar binding region to enhance the affinity, or by introducing positive charges to enhance the electrostatic interaction between the transmembrane region and the phospholipid bilayer membrane.

Provided herein are molecules or antibodies that immunospecifically binds to a surface unit or a transmembrane unit of a rabies virus glycoprotein (RABV-G). The molecules and antibodies typically including an antigen binding region of an antibody having six complementarity determining regions (CDRs). Also provided are antibody-conjugates (e.g., antibody-drug conjugates) and fusion proteins can include an antigen binding domain and a heterologous amino acid sequence. For example, chimeric antigen receptor (CAR) polypeptides including the provided antigen binding domains are also provided. In some forms, the CAR includes an extracellular antigen binding region, a spacer region, a transmembrane region, and intracellular signaling region. In some forms, the CAR includes a co-stimulatory domain. Nucleic acids encoding the molecules, antibodies, and fusions proteins such as CAR, and cells expressing the same are also provided. Pharmaceutical compositions including the provided compositions and methods of use thereof are also provided.