69 results about "Granzyme" patented technology

The present invention is a method for determining the identity of the epitopes recognized by T-cells. The method consists of expressing an encoded library of candidate epitope sequences in a recipient reporter cell capable of providing a detectable signal upon cytotoxic attack from a single cognate T-cell followed by contacting the reporter cells with T-cells of interest. The reporter cells with a single indicating cytotoxic attack from a T-cell are isolated and then analyzed by next-generation sequencing in order to identify the epitope sequences. Specifically disclosed is a method in which a library of candidate epitope-encoding nucleic acids are expressed in cells which feature a membrane-bound major histocompatibility complex (MHC) protein, said library produced by transfection of plasmids featuring both a nucleotide encoding the candidate epitope and a nucleotide encoding a FRET-based fluorescent protein cleaved by granzyme.

The invention relates to the immunoassay medical field, particularly provides a magnetic particle competition method chemiluminescence immunoassay assay kit and a preparation method thereof used for detecting hormones. The kit according to the invention comprises: 1) a calibrator; 2) magnetic particles which are coated with streptavidin; 3) hormone antigens of enzyme markers; and 4) a chemiluminescence substrate. Further, the method for preparing the kit according to the invention has the following steps: 1) pure raw materials are used to prepare the calibrator; 2) the antigens are used to coat the magnetic particles; 3) the antigens of the enzyme markers are prepared; 4) the calibrator, the chemiluminescence substrate and the antigens of the enzyme markers are packaged in a separated way; and 5) a finished product is packaged. The kit has the advantages of convenience, rapidness, sensitivity, stability, and the like.

This invention provides a non-radioactive assay to monitor and quantify the target-cell killing activities mediated by cytotoxic T lymphocytes (CTLs). This assay is predicated on the discovery that apoptosis pathway activation and, in particular, granzyme B activity, provides a measure of cytotoxic effector cell activity. In one embodiment, measurement of CTL-induced granzyme B activation in target cells is achieved through detection of the specific cleavage of fluorogenic granzyme B substrates. This assay reliably detects antigen-specific CTL killing of target cells, and provides a more sensitive, more informative and safer alternative to the standard ⁵¹Cr-release assay most often used to quantify CTL responses. The assay can be used to study CTL-mediated killing of primary host target cells of different cell lineages, and enables the study of antigen-specific cellular immune responses in real time at the single-cell level. As such, the assay can provide a valuable tool for studies of infectious disease pathogenesis and development of new vaccines and immunotherapies.

The present invention encompasses compounds of Formula (I) and pharmaceutically acceptable salts or hydrates thereof. The compounds are inhibitors of granzyme B and are useful for treating autoimmune and chronic inflammatory diseases. Pharmaceutical compositions and methods of use are also included.

The invention aims to provide an efficient amplification method of TILs serving as sources of cancerous pleural effusion. The TILs prepared with the method have the advantages of being high in multiplication speed, large in number, high in cell activity and the like. The method is mainly characterized in that by utilizing a cultivation bottle coated by laminin protein and fibronectin, the TILs are cultivated. The adsorption effect of the two kinds of protein can make cells in half-adherence state, therefore, the cells are favorable for being stimulated by various factors, and the two kinds of protein has the multiplication promoting effect on the TILs; by using a resistant OX-40 monoclone antibody, co-stimulatory signals of CD3+CD8+T cell subgroups in the TILs can be stimulated, cell activation is promoted, the expression of cell perforin and granular enzyme is improved, and the killing capacity of the TILs is enhanced. So far, the study report that the laminin protein, the fibronectin and the resistant OX-40 monoclone antibody are combined and used for preparing the TILs has not been given, and the method for increasing the cell number and improving the killing activity by adding the three factors in preparation of the TILs is put forward for the first time.

Granzyme B inhibitor compounds, compositions that include the compounds, and methods for using the compounds. The compounds of the invention have advantageous water solubility and effectively inhibit Granzyme B.

An ultraviolet (UV) disinfection unit for disinfecting catheter line connections is provided. The unit can be used to disinfect solution set and transfer set catheters used for peritoneal dialysis. The unit can comprise a concavely curved bottom surface. A tubing trough of the unit can allow for ambidextrous positioning.

A fusion protein including granzyme B, a cell penetrating peptide, a cleavage site, and a targeting moiety, a composition for cell membrane penetration comprising the fusion protein, and an anticancer composition comprising the fusion protein.

By adopting gene recombination technology to implement recombination of vascular endothelial growth factor (VEGF)2-5 exon and gran zyme B (GraB) gene and make VEGF2-5 and GraB be connected and formedinto fusion gene by means of a part of elastic connecon, and utilizing the specific combination of VEGF2-5 and VEGFR to guide GraB and make it target-act on the nescent vascular endothelial cell so as to attain the goal of target-resisting angiogenesis. Said invention uses prokaryocyte secretion type expression system to express recombinant VEGF-Gra fusion protein, and uses immobilized metal ion affinity chromatography and gel filtration method to purity the fusion protein, and uses GraB serine proteinase activity test experiment and chick embryo allantoic membrane angiogenesis anther membrane inhibition experiment to determine the bio-activity of recombinant VEGF-GraB fusion protein.

The present invention provides an enzyme-powered nanomotor, comprising a particle with a surface, an enzyme, and a heterologous molecule; characterized in that the enzyme and the heterologous molecule are discontinuously attached over the whole surface of the particle. The invention also provides the nanomotor for use in therapy, diagnosis and prognosis, in particular, for the treatment of cancer. Additionally, the invention provides the use of the nanomotor for detecting an analyte in an isolated sample.

The invention discloses an immune cell capable of simultaneously expressing a fusion protein of an intracellular truncated TGF-beta receptor and a cytokine intracellular stimulation domain and a chimeric antigen receptor on a cell membrane. The immune cell can enhance the proliferation capacity of the cell, reduce the influence of a tumor microenvironment, enhance the tumor killing activity of the CAR-T cell and reduce the depletion under the condition of receiving antigen stimulation, so that the negative regulation of the tumor microenvironment on the immune cell is weakened, the depletion of the immune cell is reduced, the tumor killing function of the immune cell is enhanced, and the inhibition of the tumor microenvironment on the immune cell is reduced; by combining with cell factor receptors, corresponding JAK and STAT signal channels are activated, the cells are induced to secrete various active substances such as IFN-gamma, perforin and granzyme, the growth of tumor cells is inhibited, the anti-tumor capability is enhanced, the killing function of immune cells is optimized, and the inhibition of a tumor microenvironment on the immune cells is reduced.

The invention discloses a natural killer cell line (NKL) modified by a human interferon (IFN)-alpha gene, which can stably excrete and express the human IFN-alpha and is established by transfecting the human IFN-alpha gene into an NKL cell. The cell line can be expressed in cells according to the RT-PCR and ELISA detection and identification, and the synthesized IFN-alpha is excreted in extracellular culture supernate. Further research indicates that the NKL cell line modified by the human IFN-alpha gene can effectively kill liver cancer cells, and such anti-tumor effect is related to the expression level improvement of the NKL-IFN-alpha cell, IFN-gamma, Granzyme, TNF-alpha and Perforin genes and proteins. Moreover, the modified NKL cell line is more sensitive to external stimulus. Therefore, the biological characteristics of the cell line are more favorable for the application of clinical adoptive immunotherapy.

The invention discloses an envelope production process for granzyme for a temperature-resistant feed. The process comprises the following steps of: mixing raw enzyme and a carrier, namely mixing the raw enzyme and the carrier to obtain paste, wherein the carrier consists of calcium carbonate, ethyl cellulose, corn starch and water, and the granzyme is prepared from the following raw materials in percentage by weight: 20 percent of calcium carbonate, 10 to 20 percent of ethyl cellulose, 20 to 30 percent of corn starch, 20 to 40 percent of raw enzyme and the balance of water; granulating; performing shot blasting; coating; and drying. By the process, the thermal stability, particularly the humidity resistance of the enzyme can be improved. Enteric polyacrylic resin latex is used as a coating material, so the use of an organic solvent can be avoided in the coating process, the production is safe, and three wastes are not generated.

The invention discloses an application of quercitrin in preparation of a human gamma delta T cell proliferator, and relates to the technical field of medicines. Quercitrin can remarkably promote proliferation of human gamma delta T cells and promote expression of perforin and granzyme B, so that the quercitrin is of certain dose dependency. The gamma delta T cell treated by the quercitrin can promote expression of p-ERK (Extracellular signal-Regulated Kinase)1/2 and p-Akt in active form and promote expression of an anti-apoptoasis protein Bcl-2, so that the gamma delta T cell is of certain dose dependency. The invention discovers the novel medical use of quercitrin and provides experimental references to development of tumor immunotherapy medicines.

The present invention provides a heat shock protein 90 (HSP90) chemiluminescence detection kit, which comprises: 1) a heat shock protein 90 (HSP90)) standard substance, wherein the heat shock protein 90 is short for HSP90; 2) magnetic particles coated with a HSP90 monoclonal antibody; 3) an enzyme-labeled HSP90 monoclonal antibody; 4) chemiluminescent substrates A and B acted by the enzyme; 5) a washing solution; and 6) a white opaque micro-pore plate. According to the present invention, with the kit, the HSP90 content in patient serum can be detected, and the important guide significance is provided for detection of non-small cell lung cancers and other tumors; and the kit has characteristics of high sensitivity, strong specificity, safety, and reliability.

The invention discloses an enhanced anti-tumor fusion protein, a preparation method and use thereof. The fusion protein includes at least two of a costimulatory molecule B7.1 that induces and activates T cells, a costimulatory molecule B7.2 that induces and activates the T cells, and a superantigen Staphylococcus aureus enterotoxin A. The fusion protein of the present invention constructs at leasttwo of B7.1, B7.2 and SEA into the fusion protein, and exerts the superimposed amplification effect. The biological activity is far more than that of a single source protein, the T cells are inducedand activated to kill tumors, the T cells secrets cytokines such as tumor necrosis factor alpha (TNF-alpha) and granzymes and induces the cancer cells as target cells to produce Fas and the like to cause cancer cells to apoptosis, The experiments have shown that each of the fusion proteins of the present invention has the significant killing effect on various cancers and solid tumors.

The invention discloses an immune cell for expressing a cytokine receptor fusion type chimeric antigen receptor and application of the immune cell. A cytokine receptor fusion type chimeric antigen receptor is expressed on a cell membrane of the immune cell, and a cytokine receptor intracellular domain is fused in an intracellular domain structure of the cytokine receptor fusion type chimeric antigen receptor. The immune cell can enhance the proliferation capacity of the cell, reduce the influence of a tumor microenvironment and enhance the tumor killing activity of the immune cell under the condition of receiving antigen stimulation, so that the negative regulation of the tumor microenvironment on the immune cell is weakened, the exhaustion of the immune cell is reduced, and corresponding JAK and STAT signal channels are activated by combining with a cytokine receptor, the cells are induced to secrete various active substances such as IFN-gamma, perforin and granzyme, so that the growth of tumor cells is inhibited, the anti-tumor capability is enhanced, and the inhibition of a tumor microenvironment on immune cells is reduced.

The invention relates to an immune cell treated by a magnetic field and application thereof. According to the invention, a medium static magnetic field with the intensity of 0.3T can promote secretionof CD8<+> T cell granzyme and cell factors of a mouse, increase the level of ATP and mitochondrial respiration and up-regulate expression of related genes Uqcrb and/ or Ndefs 6 of a mitochondrial respiratory chain. In addition, magnetic receptor candidate genes Isca1 and Cry1/ Cry2 participate in regulation and control of expression of Uqcrb and/ or Ndefs 6. An in-vivo experiment shows that the static magnetic field can inhibit growth and development of tumors by promoting secretion of the CD8<+>T cell granzyme and the cell factors. A killing ability test shows that static magnetic field treatment can enhance the killing ability of CD8<+> T cells, and the CD8<+> T cells treated by the magnetic field have a remarkable anti-tumor effect when being transfused back into a tumor grafted mouse.The invention not only discloses that the medium-intensity static magnetic field can enhance the killing ability of the CD8<+> T cells by promoting mitochondrial respiration, but also provides a novel method for enhancing the anti-tumor function of the CD8<+> T cells by a physical method.

The invention discloses a preparation method of a granzyme B activator curcumenol. It is found that eiprocurcumenol, curcumenol, physagulin A, physagulin F and withangulatin A can activate the expression of granzyme B in CIK cells and are effective granzyme B activators; the granzyme B activators of eiprocurcumenol, curcumenol, physagulin A, physagulin F and withangulatin A can significantly improve the killing capability of CIK cells on tumor cells. The invention further provides methods for preparing eiprocurcumenol, curcumenol, physagulin A, physagulin F and withangulatin A. The methods areindependent of repeated column chromatography and suitable for industrialized large-scale preparation. The technical scheme disclosed by the invention has not been disclosed in the prior art yet.