10 results about "Furin" patented technology

The application belongs to the technical field of biology and relates to an RSV pre-F mutant, a preparation method and application thereof. Compared with a wild-type F0 polypeptide, the RSV pre-F mutant satisfies the following conditions: (1) does not contain a furin enzyme cutting site fragment, a P27 polypeptide, a transmembrane domain and an intracellular domain; (2) has mutations S180C, S186C, A170C, V179C, A241C, Q279C, D486C and A490C; and (3) the C terminal of an F1 polypeptide is not connected or connected with an aggregation motif. The mutant and a trimeric conformation thereof are both highly stable, can self-assemble into a trimer without a heterologous trimeric domain, can induce high levels of binding antibodies and neutralizing antibodies after immunization, avoid high levels of non-neutralizing active binding antibodies produced when immunization is performed on the heterologous trimeric domain, and thus improve the quality of antibodies.

The application discloses a plant-expressed varicella-zoster virus gB recombinant antigen and a preparation method, product and application thereof, and belongs to the technical field of plant molecular biology and vaccine engineering. The technical problems to be solved are that the existing VZV gB protein has low heterologous expression, poor stability and insufficient antigen activity; the existing VZV vaccine has high production cost, is difficult to popularize and has a single antigen with an immune escape risk; and the plant expression system is not mature in VZV gB antigen production, and lacks a complete preparation scheme and related products. The technical solution is that the natural VZV gB protein is modified, a furin protease cleavage site is knocked out, and a H527P mutation is introduced, related nucleic acid molecules, vectors and plant source cells are constructed, high-activity antigens are obtained through culture and purification, and the high-activity antigens are applied to the preparation of VZV drugs / vaccines, and have the advantages of low cost, high safety and strong immunogenicity.

The invention provides a method for in-vitro expression and purification of recombinant human interleukin 15 in mammalian cells, which comprises the following steps: a, providing a first expression vector comprising nucleic acid encoding a fusion protein of maltose binding protein (MBP) and recombinant human IL-15, the fusion protein comprises the following elements from the 5'end to the 3 'end: 5'-MBP-Furin restriction enzyme cutting site-His tag-enterokinase restriction enzyme cutting site (EK)-rhIL-15-3 '; b, providing a second expression vector, wherein the second expression vector comprises nucleic acid for coding furin; c, co-transfecting the first expression vector and the second expression vector to mammalian cells, and performing fermentation culture to enable the cells to express recombinant human IL-15 protein; d, centrifuging to obtain fermentation supernate from the step c; e, carrying out affinity chromatography through a His tag, so as to obtain a His-EK-rhIL-15 fragment; f, the fragment is subjected to enzyme digestion with enterokinase, affinity purification is conducted again through the His tag, a flow-through substance is collected, and the flow-through substance comprises rhIL-15; and g, purifying the flow-through substance from the step f by anion exchange chromatography to obtain the purified rhIL-15 protein.

The application discloses a reconstructed respiratory syncytial virus F protein trimer, and relates to the technical field of genetic engineering vaccines.The application designs a RSV F protein trimer, the monomer of the trimer does not contain a p27 sequence, and a furin enzyme cleavage peptide segment aa110-aa136 is deleted; a GS linker is used for connecting between an F2 domain and an F1 domain; the application also designs point mutation combinations, the point mutation combinations can form inter-chain disulfide bonds, so that covalent connection of the trimer is realized, and the problem of trimer depolymerization is fundamentally solved; the trimer is stably kept in a pre-F state, and there is no post-F-like structure; an ELISA result shows that neutralizing epitopes are completely retained; a thermal acceleration experiment method is used to challenge the trimer antigen structure, and the result shows that the trimer antigen purity reaches more than 95% after being placed at 37 DEG C for 28 days, and the stability is very good; the trimer is used to prepare a vaccine, and the vaccine can meet the standard of a regulation requirement without freeze drying.

The invention provides a fusion protein which is characterized in that the fusion protein comprises the following elements from the 5'end to the 3 'end: 5'-MBP-Furin restriction enzyme cutting site-rhIL-7-3 ', and a method for in-vitro expression and purification of recombinant human interleukin 7 in mammalian cells, and the method comprises the following steps: a, providing a first expression vector, the first expression vector comprises nucleic acid for coding the fusion protein; b, providing a second expression vector, wherein the second expression vector comprises nucleic acid for coding furin; c, co-transfecting the first expression vector and the second expression vector to mammalian cells, and performing fermentation culture to enable the cells to express recombinant human IL-7 protein; d, centrifuging to obtain fermentation supernate from the step c; and e, purifying by using POROS XS and CHT chromatographic methods to obtain the rhIL-7 fragment.

The application discloses a combined biomarker combination and evaluation model for evaluating chronic low-grade inflammation of polycystic ovary syndrome, and belongs to the technical field of biomarkers. The combined biomarker combination comprises inflammation-related proteins and metabolites; the inflammation-related proteins comprise FURIN, CCL3, CCL8, CD38, NOS2, NOS3, IL18R1 and HGF; and the metabolites comprise lithocholic acid-3-O-glucuronide and taurine. The application provides a combined biomarker combination for diagnosing and evaluating the chronic inflammation state of polycystic ovary syndrome and an application method thereof. By jointly detecting specific inflammatory proteins and metabolites, the systemic chronic inflammation load of PCOS patients is quantitatively evaluated, and the combined biomarker combination has the beneficial effects of high stability, accurate discrimination, wide applicability and easy clinical transformation, and solves the problem that there is no objective evaluation method for the chronic inflammation state of PCOS.

The present results show for the first time that the HLA proteins HLA-H and HLA-J are specifically expressed at the surface of breast tumor cells (in particular Triple Negative Breast Cancer) and can therefore serve as specific targets for various immunotherapeutic agents, preferably CAR-expressing macrophages. The present invention therefore provides new immunotherapeutic molecules recognizing peptides that are specifically present in the HLA protein HLA-H or in the HLA protein HLA-J at the surface of breast cancer cells, nucleic acids encoding said molecules, immune cells expressing said molecules, as well as therapeutical uses and / or treatment methods involving them, for alleviating the symptoms of patients suffering from breast cancer, in particular from Triple Negative Breast Cancer. The invention specifically proposes to use furin-inhibited CAR-macrophages targeting extracellular peptides of the HLA-H or HLA-J proteins for treating patients suffering from breast cancer, in particular from Triple Negative Breast Cancer.

This invention relates to Furin / hydroxyl radical dual-responsive size-changing nanoparticles, their preparation, and applications. The Furin / hydroxyl radical dual-responsive size-changing nanoparticles use an AIE photosensitizer as the core and DSPE-PEG as the substrate. 2K MAL nanoparticles serve as carriers, with functional peptides coupled to their surfaces via Michael addition reactions. The core functional building block is a self-designed and custom-synthesized R1 peptide, which possesses a triple function: high specificity recognition by Furin protease, OH-responsive tyrosine oxidation cross-linking, and coupling with DSPE-PEG nanoparticles. Its simple sequence significantly enhances tumor selectivity. The peptide is covalently modified with natural peptides using a nanoprecipitation method, avoiding complex chemical modifications and exogenous orthogonal biological groups, thus mitigating the toxicity and synthesis costs of traditional systems. Under the dual conditions of Furin protease overexpression and the presence of OH, size transformation is triggered, enabling deep tumor penetration with small sizes and long-term intratumoral retention through in-situ large-size assembly, demonstrating clinical translational potential.

1.The name of the design product: ornament (Fulu Ruyi). 2.The use of the design product: for ornaments and decoration. 3.The design points of the design product: in shape. 4.The picture or photo that best shows the design points: perspective view 1. 5.The design points of the left and right views have been embodied in perspective view 1-perspective view 4, and the left and right views are omitted.