137 results about "Transmembrane Region" patented technology

The invention discloses a recombinant protein for detecting a para-tumor Yo antibody through a CBA method and application, and belongs to the technical field of biomedical engineering.The amino acid sequence of the recombinant protein sequentially comprises a secretory signal peptide, a CDR2 protein partial sequence, a CDR2L protein partial sequence, a transmembrane region and a fluorescent label, and the secretory signal peptide is human albumin signal peptide ALB; the transmembrane region is a CD8a hinge; and the fluorescent label is mCherry. A novel recombinant protein which can be stably over-expressed on a cell membrane of an eukaryotic cell is constructed by intercepting specific partial sequences of CDR2 protein and CDR2L protein and redesigning and fusing a fluorescent label by using a secretory signal peptide, a transmembrane sequence and a connecting peptide, and the recombinant protein retains respective core antigen regions of the CDR2 protein and the CDR2L protein, so that the specific partial sequences of the CDR2 protein and the CDR2L protein can be stably over-expressed on the cell membrane of the eukaryotic cell. The kit can effectively overcome the defects in the aspects of sensitivity and specificity, and when a CBA method is adopted for detection, the detection rate of the para-tumor Yo antibody can be remarkably increased, and the false positive rate is reduced, so that the requirements of clinical detection are better met.

The present invention provides ROR1 specific chimeric antigen receptors (CAR) and their therapeutic use. The CAR comprises a signal peptide, a ROR1 antigen binding domain, a hinge, a transmembrane domain, a co-stimulatory domain and an intracellular signaling domain. The modified immune cells endowed with such CARs are suitable for treating malignancies such as cancer, chronic lymphocyte leukemia (CLL), and acute lymphocytic leukemia (ALL).

The invention discloses a method and a device for detecting CAR (Chimeric Antigen Receptor) cells. The method comprises the following steps: taking a genome of a sample to be detected as a template, carrying out fluorescent quantitative PCR reaction by using a primer and a probe aiming at the sequence of a CD8 alpha transmembrane region-41 BB costimulatory molecule in a CAR gene to obtain a CT value, and calculating the copy number of the CAR gene according to the CT value and a standard curve to obtain the CAR cell distribution. Specific primers and probes are designed for CAR cells containing CD8 alpha transmembrane region-41BB costimulatory molecules, a fluorescent quantitative PCR method for detecting the copy number of CAR genes is developed, then the CAR cells are quantified, and it is proved that the method has wide applicability through multi-angle verification analysis and limitation of detection standards.

The present invention pertains to specific bioparticules at the surface of which are expressed multimeric protein. The bioparticles according to the present invention comprise an envelope consisting of a plasma membrane; and at least one type I or II transmembrane fusion protein anchored in said membrane, said fusion protein comprising successively a) a first monomer of a multimeric protein of interest, b) a coiled-coil domain or oligomerization sequence; and c) a domain for anchoring in the plasma membrane, consisting of a transmembrane segment and a cytosolic segment. Fragments a) and b) are exposed at the surface of the bioparticle, and fragment a) is bound to a second monomer of said multimeric protein by means of a bond which is not a peptide bond. The bioparticles according to the present invention can be used in therapy such as immunotherapy. The present invention also pertains to methods for producing such bioparticles.

The invention discloses a double-target chimeric antigen receptor capable of simultaneously targeting TLL1 and B7H3, a CAR-T cell and application of the double-target chimeric antigen receptor. The chimeric antigen receptor is a fusion protein which is sequentially composed of Omburt-scFv (SEQ ID NO: 1) targeting B7H3, a CD8 alpha hinge region and transmembrane region, a 4-1BB costimulatory domain, a CD3 zeta signal domain and TLL1-scFv (SEQ ID NO: 6) targeting TLL1 from an N terminal to a C terminal. The TLL1-scFv can be efficiently combined with TLL1 protein, can inhibit a TGF-beta signal channel and block prostate cancer cell migration, and is integrated into CAR of targeted B7H3, so that the double-target CAR-T cell with direct killing and immune microenvironment regulation functions is successfully constructed. The CAR-T cell has a remarkable killing effect on a prostate cancer cell line DU145, can be strongly activated after being co-cultured with a target cell and secretes a large amount of IFN-gamma and TNF-alpha, and shows high immunocompetence. The invention provides a new synergistic immunotherapy strategy for the B7H3-positive prostate cancer with the TGF-beta signal channel activated.

The application discloses a kind of multi-effect repair type collagen and its application in the technical field of synthetic biology, the application selects transdermal peptide TD-1, the coding gene of the transmembrane region of human XVII type collagen, extracellular sixteenth non-collagen region and extracellular fifteenth collagen region and His tag is connected in series, and gene sequence is optimized by pichia pastoris codon selection preference, then obtains T-COL17R3 by construction and expression. Through efficacy experiment, it is verified that T-COL17R3 compared with similar products on sale, not only excellent transdermal performance, also has more optimal ability of promoting cell proliferation, migration, anti early glycation product ketone amine and anti late glycation product dicarbonyl compound, better free radical scavenging ability and anti-elastase ability, and can reduce the synthesis of melanin in B16-F10 cell, therefore, T-COL17R3 has excellent application potential in the development of drug composition or skin care product with repair, anti-wrinkle firming or whitening effect.

The invention relates to the technical field of artificial design of enzymes, in particular to an artificial intelligence design and expression system construction method and system for lysosomal membrane protein type N-acetyltransferase with 11 transmembrane regions, and the method comprises the following steps: collecting reaction rate information of an enzyme and a substrate, associating a structure model with rate parameters, and comparing three-dimensional difference of residues, according to the method, by collecting the reaction rate correlation structure conformation, dynamic recognition of the key conformation state is achieved, the accuracy of three-dimensional space difference analysis is improved, host expression optimization factors are fused in the construction process, and the construction efficiency is improved. According to the method, the expression efficiency is improved, conformation function screening and structural stability parallel evaluation are carried out, the screening accuracy of efficient catalysis and stable expression is enhanced, three links of recognition, screening and construction are broken through, the target enzyme obtaining efficiency and expression quality are improved, and enzyme engineering is promoted to be developed towards high-throughput systematization.

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.

Provided are chimeric antigen receptors or functional fragments or variants thereof having a hinge, transmembrane region, and / or intracellular domain of LILRB1. Also provided herein are cells comprising the LILRB1-based receptors, and methods of making and using the same.

The application discloses a chimeric antigen receptor targeting CEA and a T cell containing the chimeric antigen receptor targeting CEA. The chimeric antigen receptor is composed of a nanobody recognizing a CEA antigen, an extracellular hinge region, a transmembrane region, an intracellular signal region, a self-cleavage polypeptide T2A and a hyaluronidase in sequence. The chimeric antigen receptor can efficiently recognize the CEA antigen, and CD28 and CD137 are used as a costimulatory signal region to activate T cells, thereby playing a cellular immune role, and specifically killing CEA-positive tumor cells, and thus having an important application prospect in the field of tumor cell immunotherapy.

The invention belongs to the technical field of tumor immunotherapy, and relates to an anti-Claudin18.2 single-domain antibody, a multifunctional fusion protein, a recombinant natural killer cell (CT-CAR-NK), and preparation and application thereof. Through alpaca immunization and phage display library construction and panning, the single-domain antibody VHH-C18.2-1 specifically combined with Claudin18.2 is obtained, and the amino acid sequence of the single-domain antibody VHH-C18.2-1 is SEQ ID NO: 1. The amino acid sequence of the designed fusion protein is SEQ ID NO: 3, the fusion protein sequentially comprises a VHH-C18.2-1, a flexible Linker, a TGF-beta RII extracellular domain, a CD8alpha hinge region, a CD8alpha transmembrane region, a 4-1BB intracellular domain and a CD3zeta intracellular domain from the N end to the C end, and the fusion protein has the functions of targeting, resisting TGF-beta inhibition and activating signals. The fusion protein gene transfects human peripheral blood CD56 + CD3-NK cells through lentivirus to obtain CT-CAR-NK, in-vitro verification shows that the CT-CAR-NK still keeps efficient killing in an immunosuppression environment, tumor growth can be remarkably inhibited in vivo, the lifetime can be prolonged, and a safe and efficient scheme is provided for Claudin18.2 positive solid tumor treatment.

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 present invention relates to synthetic designs or chimeric proteins for pseudotyping baculovirus (Autographa californica nuclear polyhedrosis virus) with the rabies virus glycoprotein G (gG) on its surface (Bac::gG-FL), which can be used in anti-rabies vaccine formulations. The chimeric protein is designed from a gene cassette containing gene fragments of the ectodomain of the G glycoprotein of the Pasteur strain rabies virus, a linker of 7 amino acids (GGGGSGG), as well as transmembrane (TM) and cytoplasmic (CT) regions of the gp64 baculovirus protein, with the arrangement of the sequences in the designed gene cassette being shown in figure 1.

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).

In the present invention, there is discovered a novel mRNA vaccine for preventing respiratory syncytial virus (RSV) infections, the mRNA vaccine being superior in terms of immunogenicity and pharmacological efficacy compared with RSV FG chimeric protein vaccines each comprising RSV F and G proteins and mRNA vaccines which have been produced on the basis of Japanese Patent No. 7253034 (Patent Document 7). In the present invention, an RSV FG chimeric mRNA vaccine is produced by inserting a highly conserved domain of the RSV G protein into a basic skeleton that is formed by linking a transmembrane region to an RSV F protein ectodomain. As a result of performing evaluation on immunogenicity and pharmacological efficacy, it has been confirmed that the immunogenicity and pharmacological efficacy of the RSV FG chimeric mRNA vaccine of the present invention are superior compared with those of the RSV FG chimeric protein vaccines and mRNA vaccines which have been produced on the basis of Patent Document 7.

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 invention provides a chimeric antigen receptor (CAR) targeting BCMA and GPRC5D double antigens and application of the chimeric antigen receptor. The chimeric antigen receptor (CAR) comprises an extracellular positioning signal domain, an antigen structural domain targeting BCMA, an antigen structural domain targeting GPRC5D, a hinge region, a transmembrane region, a costimulatory factor and a CD3xi intracellular signal domain, the antigen structural domain of the targeted BCMA comprises 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 structural domain of the targeted GPRC5D comprises 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 double-chimeric antigen receptor comprises an antigen structural domain targeting BCMA and an antigen structural domain targeting GPRC5D, two anti-tumor targets can be recognized at the same time, and tumor immune escape is prevented.

The invention belongs to the field of medicines, and relates to a gamma delta T cell containing a chimeric antigen receptor EGFR-CCR6-E27CAR. The chimeric antigen receptor comprises a single-chain antibody ScFv of a targeted EGFR (epidermal growth factor receptor), a CD8 hinge region, a CD8 transmembrane region, 4-1BB, a CD3 zeta chain, CCR6 and a secreting type ScFv of a targeted PD-1. The invention also comprises a protein and a nucleic acid sequence of the chimeric antigen receptor, and a preparation method and application of the vector, the protein, the nucleic acid and the cell. In an in-vitro cell model and an in-vivo mouse, the CAR-gamma delta T cell disclosed by the invention shows relatively strong tumor cell killing ability, can remarkably control in-vivo growth of tumors and remarkably prolong the lifetime of the mouse, and lays a foundation for the development of drugs for long-term control of in-vivo tumors of patients.

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.

The application discloses a recombinant pseudorabies virus strain expressing classical swine fever virus E2 protein and application, and relates to the technical field of biology.The recombinant E2 protein provided by the application successfully realizes high-level expression of the E2 protein by removing the transmembrane region of the E2 protein and using the 18aa signal peptide of the E2 protein itself and a pig albumin signal peptide.The recombinant PRV expressing the classical swine fever virus (CSFV) E2 protein can induce E2 antibodies and cellular immunity after immunization of mice.The recombinant pseudorabies virus strain provided by the application can be used as a bivalent vaccine for preventing CSFV and PRV infection.

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.

The application discloses a kind of FAP targeted chimeric antigen receptor regulatory T cells, its preparation method and application, belong to biomedical technology field.The FAP targeted chimeric antigen receptor regulatory T cell contains chimeric antigen receptor, the chimeric antigen receptor includes single-chain antibody targeted to FAP, CD8 alpha hinge region, CD8 transmembrane region, CD28 signal region and CD3 zeta signal region, its nucleotide sequence is as shown in SEQ ID NO.1.The FAP targeted chimeric antigen receptor regulatory T cell can effectively accumulate in damaged heart, and control the excessive fibrosis and inflammatory response in MI, which not only highlights the therapeutic potential of CAR Tregs against FAP in promoting heart healing, but also lays the foundation for CAR Treg treatment in clinical environment of severe MI.

Provided are an engineered G protein-coupled receptor, a complex comprising the engineered G protein-coupled receptor, and uses thereof. The modified G protein coupled receptor sequentially comprises an N terminal, a first transmembrane region, a first intracellular ring, a second transmembrane region, a first extracellular ring, a third transmembrane region, a second intracellular ring, a fourth transmembrane region, a second extracellular ring, a fifth transmembrane region, a third intracellular ring, a sixth transmembrane region, a third extracellular ring, a seventh transmembrane region and a C terminal from an N terminal to a C terminal, wherein at least a portion of the first intracellular loop, the second intracellular loop and / or the third intracellular loop is replaced with an optional first linker, a first portion of a fluorescent protein and an optional second linker, and wherein the first linker and the second linker independently comprise one or more amino acids.

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 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.

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.