339 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 preparation method comprises the following steps: firstly, analyzing the amino acid sequence of an alpha 1 chain of the human XVII type collagen, and selecting partial amino acid fragments which are not easily degraded by endogenous or exogenous protease in non-transmembrane regions of different functional domains of the protein; the recombinant collagen gene fragment (Col17a) n is obtained by repeatedly splicing the recombinant collagen gene fragment (Col17a) n; then introducing homologous promoters from different methylotrophic yeasts to the 5'end of a recombinant collagen gene segment, optimally synthesizing selected (Col17a) n and promoter segments, obtaining a connecting segment by utilizing PCR cloning, connecting the collagen segment (Col17a) n, the promoter segments and a pPIC9K carrier skeleton segment by virtue of a Gibson Assessment seamless connection technology, and obtaining the recombinant collagen gene. And constructing to obtain the pichia pastoris expression plasmid pPIC9K-(Col17a) n of the recombinant human source XVII type collagen.

The invention discloses an envelope glycoprotein for packaging a transduction NK cell pseudotyped lentiviral vector and application of the envelope glycoprotein. The envelope glycoprotein comprises an extracellular region, a transmembrane region and a cytoplasm region, wherein the amino acid sequence or the mutation sequence of the extracellular region is as shown in SEQ ID NO: 1 or SEQ ID NO: 2; an amino acid sequence or a mutation sequence of the transmembrane region is as shown in SEQ ID NO: 3 or SEQ ID NO: 4; an amino acid sequence or a mutation sequence of the cytoplasmic region is shown as SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO: 8. The BaEV envelope glycoprotein mutant replaces VSV-G to package lentivirus pseudovirus, the high virus titer 7.33 E + 07 TU / mL is maintained, the NK cells can be effectively recognized, the virus can be mediated to enter the cells, and therefore the purpose of efficiently transfecting the NK cells is achieved.

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 present invention discloses a nano antibody against BCMA and its preparation method and application, and relates to the field of biomedical technology. The present invention uses CRISPR / Cas9 technology to knock out B2M and TRAC genes in T cells to prepare universal CAR-T cells; the nano antibody targeting BCMA and the natural ligand SIRPα extracellular N-terminal V region of CD47, the transmembrane region of CD28, the intracellular signal region of 4-1BB and the intracellular signal region of CD3ζ are connected in series to form a dual-targeted chimeric antigen receptor for the treatment of multiple myeloma, so as to solve the high recurrence problem faced by CAR-T cell therapy in the treatment of multiple myeloma. The universal CAR-T cells prepared by the present invention have strong killing activity against multiple myeloma cells, effectively improve the treatment effect, and provide a key material for the subsequent development of adoptive immunotherapy with high efficiency and lasting anti-tumor activity.

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.

The invention provides a CAR treatment method for treating acute myelogenous leukemia and application of the CAR treatment method. The invention optimizes and establishes a chimeric antigen receptor (CAR) aiming at acute myelogenous leukemia (AML). The chimeric antigen receptor comprises an extracellular binding region, a transmembrane region, an intracellular signal region and an X molecule which are connected in sequence. The invention also provides an immune effector cell modified by the CAR. The invention also discloses a preparation method of the compound and application of the compound in inhibition of acute myelogenous leukemia.

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 present invention relates to a molecular switch regulation type chimeric antigen receptor polypeptide, containing a humanized anti-P329G mutant scFv sequence, a hinge region / spacer region, a transmembrane region, a costimulatory signal domain, and a stimulation signal domain; an immune effector cell engineered to express the molecular switch regulation type chimeric antigen receptor polypeptide; and a method for preparing the immune effector cell. The present invention further relates to a P329G mutant antibody capable of specifically binding CLDN18.2 molecules and a method for preparing the P329G mutant antibody. A pharmaceutical combination containing the immune effector cell and the P329G mutant antibody can be used for treating diseases related to CLDN18.2, such as cancer expressing or overexpressing CLDN18.2.

The invention discloses a chimeric antigen receptor simultaneously targeting GPRC5D and CS1 and application thereof, and relates to the technical field of biological medicines. Spatial epitopes of a GPRC5D antigen and a CS1 antigen are taken as target spots, an scfv region of a GPRC5D antibody and an scfv region of a CS1 antibody are connected in series through Linker, and an optimal structure VH1-VL1-VH2-VL2 is obtained through screening, VH1 and VL1 are anti-GPRC5D scfv, and VH2 and VL2 are anti-CS1scfv. The chimeric antigen receptor constructed by connecting the optimal structure VH1-VL1-VH2-VL2 with a transmembrane region of CD8, a CD137 intracellular signal region and a CD3zeta intracellular signal region in series is used for treating the multiple myeloma, so that the problems of low remission rate and high relapse rate of a CAR-T cell therapy in treatment of the multiple myeloma are solved, and the chimeric antigen receptor is still effective for the multiple myeloma with BCMA negative or low BCMA antigen expression and can be used for treating the multiple myeloma. And a new strategy is provided for clinical treatment of multiple myeloma patients with negative BCMA expression or recurrence after BCMA CAR-T treatment.

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

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.

The invention relates to application of anti-PTPRZ1 chimeric antigen receptor macrophages in lung cancer treatment, and belongs to the technical field of tumor targeted therapy. The invention constructs a novel chimeric antigen receptor (CAR) which comprises a coding sequence of a single-chain variable region fragment for specifically recognizing PTPRZ1, and a coding sequence of a CD8TM transmembrane region and a coding sequence of a CD3zeta intracellular signal transduction structural domain. The design ingeniously endows the chimeric antigen receptor (CAR) with high specificity, so that the chimeric antigen receptor (CAR) can accurately recognize and firmly bind to the surface of a tumor cell expressing PTPRZ1 protein. According to the present invention, the chimeric antigen receptor targeting PTPRZ1 is successfully transfected into the human macrophage to form the CAR-M. A flow cytometry verifies that compared with non-transfected macrophages (Mphi) or transfected macrophages lacking a specific target binding domain (CAR-M-delta), the CAR-M has the advantage that the efficiency of swallowing PTPRZ1 positive tumor cells is remarkably improved in the aspect of the efficiency of swallowing PTPRZ1 positive tumor cells. Therefore, the CAR-M constructed by the invention can be used as a potential treatment means for targeting PTPRZ1 expression tumors.

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 enhanced chimeric antigen receptor and application thereof, in particular to a nucleic acid molecule encoding a chimeric antigen receptor containing an NKG2D transmembrane region, a corresponding chimeric antigen receptor, a carrier, an immune effector cell, a preparation method and a product thereof, a pharmaceutical composition, pharmaceutical application and a tumor or cancer treatment method. The CAR has the advantages of high CAR expression efficiency, high NK cell proliferation speed, high killing power to tumor cells, high specificity, long killing effect duration and the like.

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.

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

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 discloses an envelope glycoprotein for packaging a transduction NK cell pseudotyped lentiviral vector and application of the envelope glycoprotein. The envelope glycoprotein comprises an extracellular region, a transmembrane region and a cytoplasm region, wherein the amino acid sequence or the mutation sequence of the extracellular region is as shown in SEQ ID NO: 1 or SEQ ID NO: 2; an amino acid sequence or a mutation sequence of the transmembrane region is as shown in SEQ ID NO: 3 or SEQ ID NO: 4; an amino acid sequence or a mutation sequence of the cytoplasmic region is shown as SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO: 8. The BaEV envelope glycoprotein mutant replaces VSV-G to package lentivirus pseudovirus, the high virus titer 7.33 E + 07 TU / mL is maintained, the NK cells can be effectively recognized, the virus can be mediated to enter the cells, and therefore the purpose of efficiently transfecting the NK cells is achieved.

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 invention discloses a chimeric antigen receptor double-negative T cell drug co-expressing CXCR5 and IL-7 and application thereof, the cell drug can co-express CXCR5 and IL-7, and the cell drug comprises EF1-alpha, a CD8 alpha signal peptide, an NKG2D extracellular domain, a CD8 alpha hinge region, a CD8 alpha transmembrane region, a costimulatory factor 4-1BB, a CD3 zeta intracellular signal region, T2A, a chemotactic factor receptor CXCR5, P2A and interleukin IL-7 which are connected in sequence; according to the chimeric antigen receptor double-negative T cell drug co-expressing the CXCR5 and the IL-7 and the application of the chimeric antigen receptor double-negative T cell drug, the CXCR5 and the IL-7 are co-expressed on CAR-DNT cells of targeted NKG2DL, so that the migration and infiltration capacity of the drug to a tumor site is enhanced, the survival and proliferation capacity of the CAR-T cells is improved, the killing effect of the CAR-DNT cells on tumors is improved, and the drug has a good application prospect. The purpose of treating tumors is achieved, and allogenic treatment can be achieved.

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).