249 results about "Transmembrane Region" patented technology

The invention relates to a recombinant A type humanized XVII type collagen as well as construction and application thereof, the protein is the humanized XVII type collagen which is composed of an intracellular region, a transmembrane region and an extracellular region, and the amino acid sequence is as shown in SEQ No. 1. The hair follicle repairing agent has the advantages of hair follicle repairing, hair regeneration, good biological activities of cell proliferation, cell adhesion and the like, and can be widely applied to the fields of medicines, medical instruments, biological materials, tissue engineering, cosmetics and the like. The gene sequence of the recombinant A type XVII collagen is 100% identical to the corresponding part of the gene sequence of the recombinant A type XVII collagen, so that the recombinant A type XVII collagen has good hydrophilicity and high activity, and has application potential in the fields of biomedical materials, beauty cosmetics, skin care products and the like.

The invention discloses a multi-effect repair type collagen and application thereof, and belongs to the technical field of synthetic biology. Transdermal peptide TD-1, a transmembrane region of human XVII type collagen, an extracellular sixteenth non-collagen region, an extracellular fifteenth collagen region and a coding gene of a His tag are selected to be connected in series, and a gene sequence is optimized through pichia pastoris codon selection preference; and then the T-COL17R3 is constructed and expressed to obtain the T-COL17R3. Efficacy experiments prove that compared with similar products sold in the market, T-COL17R3 not only has excellent transdermal performance, but also has better cell proliferation and migration promoting capability, better capability of resisting early saccharification product ketoamine and later saccharification product dicarbonyl compounds, better capability of scavenging free radicals and resisting elastase, and can be used for preparing the anti-epidermal drug. In addition, synthesis of melanin in B16-F10 cells can be reduced, so that T-COL17R3 has excellent application potential in development of pharmaceutical compositions or skin care products with repairing, anti-wrinkle and firming or whitening and brightening effects.

The invention belongs to the technical field of immunology and in vitro diagnosis, and relates to a monoclonal antibody aiming at bovine nodular skin disease virus (LSDV) L1R protein. According to the invention, the LSDV recombinant L1R protein with deletion of a transmembrane region (182-204aa) is obtained through expression; after a mouse is immunized by the protein, a monoclonal antibody 8D5 which is good in reactivity and specificity and aims at the LSDV L1R protein is screened out; heavy chain and light chain variable region sequences for coding the monoclonal antibody 8D5 are obtained; the monoclonal antibody 8D5 can be subjected to specific reaction with LSDV L1R protein, the recombinant protein L1R can be specifically recognized by bovine LSDV positive serum, meanwhile, the monoclonal antibody 8D5 can be used for establishing an LSDV ELISA antibody detection method, and a foundation is laid for LSDV diagnostic technology and vaccine research and development.

The invention provides an establishment method of a transgenic mouse, and the method comprises the following steps: a transgenic mouse MHC chimeric gene is introduced into an endogenous B2m gene locus of the mouse, the MHC chimeric gene comprises a human B2M gene and a chimeric alpha-chain gene, and the chimeric alpha-chain gene comprises a human alpha1 region, a human alpha2 region and a mouse alpha3 region; the chimeric alpha-chain gene further comprises a mouse transmembrane region and a mouse intracellular region. The transgenic mouse disclosed by the invention can be applied to tumor specific antibody detection, in-vivo evaluation of vaccines, and in-vivo screening of TCR-T and other drugs.

The invention belongs to the technical field of biological medicine, and particularly relates to a preparation method of viroid particles displaying scFv antibodies, a product of the viroid particles and application of the viroid particles. The preparation method provided by the invention comprises the following steps: constructing a specific targeting recombinant plasmid, wherein the specific targeting recombinant plasmid is obtained by connecting a vector for expressing virus capsid protein and a coding gene for expressing an scFv antibody display structure through homologous recombination. The vector for expressing the virus capsid protein comprises a coding gene of VSV-G signal peptide, a transmembrane region and an intracellular region, and is as shown in SEQ ID NO. 2. And the coding gene for expressing the scFv antibody display structure is shown as SEQ ID NO. 3-5. According to the preparation method disclosed by the invention, the virus-like particles of which the capsid is a modular component of a specific targeting immune cell are constructed based on a lentivirus system, and 50-90% of infection efficiency can be realized in the immune cell.

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 invention discloses an E-CAR-NK cell for treating systemic lupus erythematosus and application of the E-CAR-NK cell, and belongs to the technical field of genetic engineering.A CD19 and BCMA double-target E-CAR is adopted for modifying an NK cell, and the E-CAR-NK cell is obtained; the CD19 and BCMA double-target E-CAR is composed of the following modules: a CD8 signal peptide, a CD19 scFv, a Linker, a BCMA scFv, a CD8 Hinge region, an NKG2D transmembrane region, a CD3 epsilon signal transduction region, a DAP12 costimulatory region, a 2B4 costimulatory region, and a CD3 zeta signal transduction region. Compared with a common CAR-NK cell, the E-CAR-NK cell disclosed by the invention has higher antigen sensitivity and better continuous killing ability, so that the treatment effect of SLE is improved, and the E-CAR-NK cell has lower side effects and higher safety.

The invention discloses an MUC1-CAR-T cell for co-expressing a BTLA / IL-18R chimeric receptor as well as a preparation method and application of the MUC1-CAR-T cell, and belongs to the technical field of biological medicines. The MUC1-CAR-T cell comprises a BTLA / IL-18R chimeric receptor and an MUC1-CAR structure, wherein the BTLA / IL-18R chimeric receptor is a BTLA MUC1 is used as a targeting molecule for CAR-T cell therapy, an extracellular domain of BTLA is connected with a transmembrane domain and an intracellular domain of an IL-18 receptor, and the co-expression BTLA / IL-18R chimeric receptor is designed. Experimental verification shows that the BTLA / IL-18R chimeric receptor can significantly improve the tumor killing ability of MUC1-CAR-T cells, can better inhibit the growth of tumors, and provides a novel immunotherapy strategy for the treatment of pancreatic cancer.

Provided is a chimeric antigen receptor targeting GCC, comprising: an scFv that specifically recognizes GCC, a CD8 hinge region or a CD28 hinge region, a CD8 transmembrane region or a CD28 transmembrane region, a CD28 co-stimulatory signal domain or a 4-1BB co-stimulatory signal domain, and a CD3ζ signal domain; the scFv that specifically recognizes GCC comprises a heavy chain variable region VH and a light chain variable region VL, the VH comprising an HC CDR1 having the amino acid sequence shown in SEQ ID NO: 1, an HC CDR2 having the amino acid sequence shown in SEQ ID NO: 2, and an HC CDR3 having the amino acid sequence shown in SEQ ID NO: 3, and the VL comprising an LC CDR1 having the amino acid sequence shown in SEQ ID NO: 4, an LC CDR2 having the amino acid sequence shown in SEQ ID NO: 5, and an LC CDR3 having the amino acid sequence shown in SEQ ID NO: 6.

The invention relates to the technical field of biomedicine, in particular to a fusion protein combining a double-target chimeric antigen receptor with PD-1 and IL-21, and an extracellular domain of the double-target chimeric antigen receptor sequentially comprises any one of the following components from an N terminal to a C terminal: CIgG scFv-MSLN scFv, CIgG scFv-CLDN18.2 scFv, CIgG scFv-MSLN scFv and CIgG scFv-CLDN18.2 scFv; wherein '-' is a connecting peptide; the cell further comprises a hinge region, a transmembrane region and an intracellular signal transduction region; pD-1 scFv and IL-21 are further fused at the N end of the intracellular signal transduction region. According to the invention, two chimeric antigen receptors of double-target combined PD-1 and IL21 fusion proteins are constructed, and through a synergistic anti-tumor strategy, CAR-T cell functions are enhanced, tumor microenvironment (TME) inhibition is overcome, T cell durability is improved, and synergy is realized.

The invention discloses a VZV antigen variant, nucleic acid, a pharmaceutical composition and application of the VZV antigen variant. The VZV antigen variant has a Y582G difference compared with an amino acid sequence as shown in SEQ ID NO: 1; and / or the VZV antigen variant has the deletion of 561 to 623 sites, 569 to 623 sites or 574 to 623 sites compared with the amino acid sequence as shown in SEQ ID NO: 1, and / or the VZV antigen variant has the deletion of 561 to 623 sites, 569 to 623 sites or 574 to 623 sites; and / or the VZV antigen variant is a modification having a protein transmembrane region and an intracellular region compared with the amino acid sequence as shown in SEQ ID NO: 1, and the modification of the protein transmembrane region and the intracellular region is that the transmembrane region and the intracellular region of the original VZV antigen are replaced by a transmembrane region of SARS-CoV-2Spike protein or a transmembrane region of influenza H protein. Compared with the VZV antigen in the prior art, the VZV antigen variant has stronger immunogenicity, can obtain higher binding antibody titer, and has higher protective efficacy.

The invention relates to an IL5RA cell surface marker. A cell tag comprising an extracellular region, a transmembrane region, and optionally an intracellular region is provided. The extracellular region comprises an IL5 receptor alpha (IL5Ra) sequence linked to a transmembrane domain, and the recombinant polypeptide cannot function in signal transduction. The cell tag can be operably linked to a transgene. Expression of the cell tag allows for the identification, detection, selection, and ablation of cells expressing the transgene and the cell tag. In some embodiments, the genetically modified host cell comprises a transgene comprising a polynucleotide encoding a chimeric antigen receptor comprising a ligand binding domain and a polynucleotide encoding a cell tag. Pharmaceutical formulations produced by the methods and methods of using the pharmaceutical formulations are also described.

A method for embedding a nanopore protein into a biomimetic membrane and the use thereof. The method comprises: preparing a biomimetic membrane of a multi-block copolymer, wherein the multi-block copolymer comprises a hydrophilic segment and a hydrophobic segment; and embedding a nanopore protein into the biomimetic membrane, wherein the multi-block copolymer comprises a copolymer of two or more blocks, the biomimetic membrane comprises an aromatic ring structure, the aromatic ring structure is linked between the hydrophilic segment and the hydrophobic segment of the multi-block copolymer, and / or the nanopore protein comprises an aromatic ring structure, and the aromatic ring structure is located on an amino acid in the transmembrane region of the nanopore protein facing the biomimetic membrane. By means of the method, the number of single pores and / or the single-pore retention rate of a biomimetic membrane during embedding can be improved.

Provided is a genetically engineered cell, which expresses a co-receptor and a membrane surface cytokine. The co-receptor contains a KIR extracellular domain, a transmembrane region and an intracellular region. When the co-receptor and the membrane surface cytokine are co-expressed in an immune cell, the cytokine secretion level, proliferation level and killing ability for a target cell of the immune cell can be further improved.

The invention relates to an RSV F protein soluble fragment, an autotrimer structural domain and application thereof, and belongs to the technical field of recombinant protein. Based on an amino acid sequence of a wild RSV F protein, a soluble and rigid sequence is screened out, a soluble fragment is constructed, disulfide bond mutation pair modification is performed on the basis of the soluble fragment to obtain a self-derived trimer structural domain sequence, and the self-derived trimer structural domain sequence is used for replacing a transmembrane region and an intracellular region of the RSV F protein. Therefore, trimerization can be realized, stable pre-fusion conformation can be maintained, and soluble expression in cells can be realized at the same time. The invention also provides an RSV F protein mutant containing an autotrimer structural domain sequence, the generation and the stability of the pre-F conformation trimer are proved, a representative is selected from the pre-F conformation trimer for animal experiments, and it is proved that the pre-F conformation trimer has good immunogenicity and can be used for developing RSV vaccines and preparing RSV specific antibodies or RSV detection reagents.

The present invention provides a combined chimeric antigen receptor targeting CD19 and CD20 and application thereof. Specifically, the present invention provides a combined chimeric antigen receptor targeting CD19 and CD20, which comprises a scFv targeting CD19 and CD20, a hinge region, a transmembrane region, and an intracellular signaling domain. The present invention provides a nucleic acid molecule encoding the chimeric antigen receptor and a corresponding expression vector, a CAR-T cell, and applications thereof. The experimental results show that the chimeric antigen receptor provided by the present invention shows extremely high killing ability against tumor cells. The chimeric antigen receptor of the present invention targets CD19 and / or CD20 positive cells and can be used to treat CD19 and / or CD20 positive B-cell lymphoma, leukemia and other diseases.

The invention provides a CLL1-targeting enhanced chimeric antigen receptor and application thereof. The CLL1-targeting enhanced chimeric antigen receptor comprises a CLL1-targeting extracellular antigen recognition structural domain, a hinge region, a transmembrane region and an intracellular structural domain, and the intracellular structural domain comprises a costimulatory factor and an intracellular signal transduction structural domain; wherein the intracellular domain further comprises a cytokine receptor gamma chain amino acid sequence derived from IL2R gamma, or wherein the costimulatory domain comprises an amino acid sequence derived from 2B4. Compared with a CLL1CAR-DNT cell with a conventional second-generation chimeric antigen receptor structure, the CLL1CAR-DNT cell with the enhanced chimeric antigen receptor structure has the advantage that the proliferation level of the CLL1CAR-DNT cell with the enhanced chimeric antigen receptor structure is higher.

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 provides a membrane-wrapped organelle-like system for directionally assembling membrane protein, which comprises a molecular framework and a closed lipid bilayer membrane wrapping the outer side of the molecular framework, the molecular framework is loaded with load molecules, the molecular framework directionally recombines the lipid bilayer membrane through a transmembrane region of the membrane protein, and the lipid bilayer membrane is loaded with the load molecules. The lipid bilayer membrane provides a barrier and protection for a molecular framework and loaded molecules, and the lipid bilayer membrane is provided with directional recognition molecules. According to the invention, a molecular framework wrapped by a lipid bilayer membrane is utilized, and a novel technology capable of simulating lipid vesicles of an organelle membrane framework is adopted, so that a brand-new directional delivery function on specific cells or specific organelles in the specific cells can be endowed. And oriented assembly of a membrane protein compound and effective loading of bioactive molecules are realized by using a process of chemically recombining the lipid bilayer membrane.

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.

Provided herein, amongst other things, are polynucleotides comprising a) a chimeric antigen receptor (CAR) comprising: an extracellular antigen binding domain comprising an antibody or antigen binding fragment thereof; a transmembrane region; and an intracellular signaling region comprising an OX40L intracellular signaling domain; and b) a nucleic acid encoding an IL-15.

The present invention provides a GD2-targeting CAR-T cell and its preparation and application. Specifically, the present invention provides a GD2-targeting CAR construct, which comprises the scFv fragment of the humanized GD2 antibody 3F8, the human Fc fragment, the ICOS transmembrane region, the ICOS and 4-1BB intracellular regions (co-stimulatory signals), and CD3ζ. The present invention also provides a CAR-T cell based on the above CAR, its preparation method and application. The CAR-T cell of the present invention can significantly inhibit the proliferation of tumor cells in a subcutaneous model of neuroblastoma, has a significant in vivo anti-tumor effect, and can be used for the targeted treatment of neuroblastoma.

The present disclosure provides a chimeric polypeptide. The chimeric polypeptide includes: an extracellular region having an activity of binding to a first molecule; a transmembrane region having an N-terminus linked to a C-terminus of the extracellular region; and an intracellular region having an N-terminus linked to a C-terminus of the transmembrane region, wherein the transmembrane region includes an isolated polypeptide having at least 80% identity with a transmembrane region of a Notch receptor protein derived from Xenopus tropicalis.

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.