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1
EP2899269B1
Publication/Patent Number: EP2899269B1
Publication date: 2019-04-17
Application number: 13823838.1
Filing date: 2013-07-24
Abstract: An object is to provide a TCR cloning system that enables not only bias-free analysis of TCR repertoires, but also collection of antigen-specific TCR α/β cDNA pairs and evaluation of functions thereof. There is provided a method for producing a gene of T cell receptor (TCR) specific to an antigen A, which comprises 1) the step of stimulating a group of T cells including a T cell specific to an antigen A or one T cell specific to an antigen A under a condition effective for amplification of a TCR gene; 2) the step of identifying a T cell specific to an antigen A among the group of T cells including a T cell specific to the antigen A, and sorting one T cell specific to the antigen A into a vessel; and 3) the step of subjecting the one activated T cell specific to the antigen A in the vessel to PCR to amplify a gene of TCR specific to the antigen A. According to the present invention, a target TCR gene can be cloned within a shorter time compared with that required by the conventional methods, for example, about ten days. Further, according to the present invention, genes of TCR α chain and β chain can be highly efficiently cloned. Under the conditions of the examples, a pair of a TCR α chain and TCR β chain could be obtained from stimulated T cells sorted as single cells at a ratio of 100%. An object is to provide a TCR cloning system that enables not only bias-free analysis of TCR repertoires, but also collection of antigen-specific TCR α/β cDNA pairs and evaluation of functions thereof. There is provided a method for producing a gene of T cell receptor (TCR) s ...more ...less
3
EP2824182B1
Publication/Patent Number: EP2824182B1
Publication date: 2019-01-02
Application number: 13758671.5
Filing date: 2013-03-06
Abstract: An object of the present invention is to stimulate a T cell without using a peptide/MHC tetramer. In the present invention, the step of supplying an antigen peptide to a T cell having a T cell receptor (TCR) that can recognize the antigen peptide on cell surface to form a complex of a major histocompatibility complex (MHC) molecule on the cell surface of the T cell and the antigen peptide is used, and the T cell is stimulated through recognition by TCR of the antigen peptide as the MHC molecule-antigen peptide complex on the cell surface of the same T cell. Such a stimulating and activating method would be applicable to not only T cells, but also various cells. According to the present invention, an antigen-specific T cell can be identified without establishing any antigen-specific T cell strain, and without using such a reagent as MHC/peptide tetramer. That is, a cancer-specific T cell can be efficiently and conveniently identified. An object of the present invention is to stimulate a T cell without using a peptide/MHC tetramer. In the present invention, the step of supplying an antigen peptide to a T cell having a T cell receptor (TCR) that can recognize the antigen peptide on cell surface to form a ...more ...less
4
CN107922912A
Publication/Patent Number: CN107922912A
Publication date: 2018-04-17
Application number: 201680046633
Filing date: 2016-08-10
Abstract: The present invention relates to a method for separating cells capable of producing target antigen-specific monoclonal antibodies. In said method: a cell group including antibody-producing cells is immobilized using a crosslinking agent (the crosslinking agent is a reversible crosslinking agent having cell membrane-permeating properties); the immobilized cell group is subjected to cell membrance dissolution using a surface active agent; and the cell group is reacted with a labeling target antigen, and in the stained cell group, at least one cell that has reacted with the labeling target antigen is separated. The present invention further relates to a method for producing target antigen-specific monoclonal antibodies. In said method: mRNA is separated from the cell separated using the aforementioned method; cDNA is prepared; and antigen-specific monoclonal antibodies or fragments thereof are prepared from the prepared cDNA. Also provided are a method with which at least one cell capableof producing target antigen-specific monoclonal antibodies can be separated with high accuracy, and a method with which target antigen-specific monoclonal antibodies can be produced by using the cellseparated using said method. The present invention further provides new threonine 18 phosphorylated p53 (pT18-p53) specific monoclonal antibodies, and new threonine 68 phosphorylated CHK2 (pT68-CHK2)specific monoclonal antibodies. The present invention relates to a method for separating cells capable of producing target antigen-specific monoclonal antibodies. In said method: a cell group including antibody-producing cells is immobilized using a crosslinking agent (the crosslinking agent is a reversible ...more ...less
5
EP2692864B1
Publication/Patent Number: EP2692864B1
Publication date: 2018-09-12
Application number: 12765903.5
Filing date: 2012-03-27
Abstract: The purpose of the invention is to provide means with which it is possible to efficiently select a vector to which a foreign gene has been introduced when a foreign gene is to be introduced by homologous recombination to a vector having multiple sequences homologous with one another. The vector comprises, in succession, a replication origin, a sequence A, a marker gene X, two sequences C and D for introducing a foreign gene by homologous recombination, and a sequence B homologous with sequence A. The two sequences C and D are directly or indirectly adjacent to one another. The vector is used for introducing a foreign gene between the two adjacent sequences C and D. The purpose of the invention is to provide means with which it is possible to efficiently select a vector to which a foreign gene has been introduced when a foreign gene is to be introduced by homologous recombination to a vector having multiple sequences homologous with one ...more ...less
6
EP3336172A1
Publication/Patent Number: EP3336172A1
Publication date: 2018-06-20
Application number: 16835229.2
Filing date: 2016-08-10
Abstract: The present invention relates to a method for separating cells capable of producing target antigen-specific monoclonal antibodies. In said method: a cell group including antibody-producing cells is immobilized using a crosslinking agent (the crosslinking agent is a reversible crosslinking agent having cell membrane-permeating properties); the immobilized cell group is subjected to cell membrance dissolution using a surface active agent; and the cell group is reacted with a labeling target antigen, and in the stained cell group, at least one cell that has reacted with the labeling target antigen is separated. The present invention further relates to a method for producing target antigen-specific monoclonal antibodies. In said method: mRNA is separated from the cell separated using the aforementioned method; cDNA is prepared; and antigen-specific monoclonal antibodies or fragments thereof are prepared from the prepared cDNA. Also provided are a method with which at least one cell capable of producing target antigen-specific monoclonal antibodies can be separated with high accuracy, and a method with which target antigen-specific monoclonal antibodies can be produced by using the cell separated using said method. The present invention further provides new threonine 18 phosphorylated p53 (pT18-p53) specific monoclonal antibodies, and new threonine 68 phosphorylated CHK2 (pT68-CHK2) specific monoclonal antibodies. The present invention relates to a method for separating cells capable of producing target antigen-specific monoclonal antibodies. In said method: a cell group including antibody-producing cells is immobilized using a crosslinking agent (the crosslinking agent is a reversible ...more ...less
8
US10023943B2
Publication/Patent Number: US10023943B2
Publication date: 2018-07-17
Application number: 15/092,934
Filing date: 2016-04-07
Abstract: An Al—Mg—Si-based aluminum alloy includes 0.015 to 0.12 mass % of Sr, the aluminum alloy producing a cast metal structure in which Mg2Si is crystallized in a fine agglomerate form.
9
EP2990035B1
Publication/Patent Number: EP2990035B1
Publication date: 2018-03-07
Application number: 14786113.2
Filing date: 2014-03-26
Abstract: A collagen production promoter in cells, containing at least one member selected from the group consisting of 1,5-anhydro-D-glucitol and derivatives thereof; and a composition containing the collagen production promoter. Since the collagen production promoter containing 1,5-AG or derivatives thereof of the present invention is suitably used as cosmetics, medicinal formulations, foods, and the like, for promoting collagen production in cells, for example, preventing and/or improving wrinkles of skin. A collagen production promoter in cells, containing at least one member selected from the group consisting of 1,5-anhydro-D-glucitol and derivatives thereof; and a composition containing the collagen production promoter. Since the collagen production promoter containing 1,5-AG ...more ...less
10
US2018057860A1
Publication/Patent Number: US2018057860A1
Publication date: 2018-03-01
Application number: 20/171,568
Filing date: 2017-08-28
Abstract: Disclosed is a thermostable DNA polymerase preparation which can illimitably reduce the risk of false positivity in the detection of a subject microorganism utilizing a gene amplification reaction and therefore enables the selective amplification of DNA for detecting the subject microorganism even when the amount of the subject microorganism is small and therefore the amount of DNA collected therefrom is extremely small, and can be produced at a reduced cost. Also disclosed is a method for quantifying or quantifying/identifying a subject organism to be detected rapidly, conveniently and with high sensitivity using the preparation of the present invention. Disclosed is a thermostable DNA polymerase preparation which can illimitably reduce the risk of false positivity in the detection of a subject microorganism utilizing a gene amplification reaction and therefore enables the selective amplification of DNA for detecting the subject ...more ...less
11
US20180292407A1
Publication/Patent Number: US20180292407A1
Publication date: 2018-10-11
Application number: 15/751,792
Filing date: 2016-08-10
Abstract: A method for separating cells capable of producing target antigen-specific monoclonal antibodies (TASMAs) wherein a cell group including antibody-producing cells is immobilized using a reversible crosslinking agent having cell membrane-permeating properties. The immobilized cell group is subjected to cell membrane dissolution using a surface active agent; and the cell group is reacted with a labeling target antigen. In the stained cell group a that has reacted with the labeling target antigen is separated. A method to produce TASMAs by separating mRNA from the cell separated using the method; preparing cDNA and preparing antigen-specific monoclonal antibodies or fragments thereof from the prepared cDNA. Also provided are a method whereby at least one cell capable of producing TASMAs is separated and a method whereby said antibodies can be produced by using the separated cell. Threonine 18 phosphorylated p53 (pT18-p53) and threonine 68 phosphorylated CHK2 (pT68-CHK2) specific monoclonal antibodies are also disclosed. A method for separating cells capable of producing target antigen-specific monoclonal antibodies (TASMAs) wherein a cell group including antibody-producing cells is immobilized using a reversible crosslinking agent having cell membrane-permeating properties. The immobilized cell ...more ...less
12
US20180057860A1
Publication/Patent Number: US20180057860A1
Publication date: 2018-03-01
Application number: 15/688,473
Filing date: 2017-08-28
Abstract: Disclosed is a thermostable DNA polymerase preparation which can illimitably reduce the risk of false positivity in the detection of a subject microorganism utilizing a gene amplification reaction and therefore enables the selective amplification of DNA for detecting the subject microorganism even when the amount of the subject microorganism is small and therefore the amount of DNA collected therefrom is extremely small, and can be produced at a reduced cost. Also disclosed is a method for quantifying or quantifying/identifying a subject organism to be detected rapidly, conveniently and with high sensitivity using the preparation of the present invention. Disclosed is a thermostable DNA polymerase preparation which can illimitably reduce the risk of false positivity in the detection of a subject microorganism utilizing a gene amplification reaction and therefore enables the selective amplification of DNA for detecting the subject ...more ...less
13
EP2412377B1
Publication/Patent Number: EP2412377B1
Publication date: 2018-12-26
Application number: 10755762.1
Filing date: 2010-02-05
Abstract: PROBLEM TO BE SOLVED A burdock fruit extract containing arctigenin at high content and its production method are provided, and both of which are used for treatment of pancreatic cancer. SOLUTION The burdock fruit extract containing arctigenin at high content by enzymatically converted arctiin into arctigenin with beta-glucosidase, which is an enzyme occurring endogenously in a burdock fruit, and adding ethanol, extracting, concentrating and then freeze-drying or spray drying. PROBLEM TO BE SOLVED A burdock fruit extract containing arctigenin at high content and its production method are provided, and both of which are used for treatment of pancreatic cancer. SOLUTION The burdock fruit extract containing arctigenin at high content by enzymatically ...more ...less
14
WO2018012602A1
Publication/Patent Number: WO2018012602A1
Publication date: 2018-01-18
Application number: 2017025598
Filing date: 2017-07-13
Abstract: A magnesium alloy containing 5.0 mass% to 15.0 mass% of Al
15
US9907852B2
Publication/Patent Number: US9907852B2
Publication date: 2018-03-06
Application number: 15/302,863
Filing date: 2015-04-10
Abstract: The purpose of the present invention is to provide an anticancer agent for potentiating an antitumor effect, alleviating side effects, and further extending the survival rate by concomitant use with a component having an anticancer effect. An anticancer agent combining arctigenin and a component other than arctigenin that has an anticancer effect, in which the anticancer agent may be a combination drug or may be a kit configured from a formulation containing arctigenin and a formulation containing a component that has an anticancer effect, and the concomitant use of arctigenin and the component having an anticancer effect more strongly inhibits tumor growth and reduces the proportion of cancer stem cells in the tumor, making it possible to extend the total survival time and to alleviate side effects caused by the component having an anticancer effect. The purpose of the present invention is to provide an anticancer agent for potentiating an antitumor effect, alleviating side effects, and further extending the survival rate by concomitant use with a component having an anticancer effect. An anticancer agent combining ...more ...less
16
US20180371583A1
Publication/Patent Number: US20180371583A1
Publication date: 2018-12-27
Application number: 15/779,971
Filing date: 2017-07-13
Abstract: A magnesium alloy is provided that includes: 5.0 mass % or more and 15.0 mass % or less of Al; 2.5 mass % or more and 7.0 mass % or less of Sr; 0.05 mass % or more and less than 3.0 mass % of Ca; and 0.1 mass % or more and 0.6 mass % or less of Mn, with a remainder including Mg and inevitable impurities. A magnesium alloy is provided that includes: 5.0 mass % or more and 15.0 mass % or less of Al; 2.5 mass % or more and 7.0 mass % or less of Sr; 0.05 mass % or more and less than 3.0 mass % of Ca; and 0.1 mass % or more and 0.6 mass % or less of Mn, with a remainder including Mg ...more ...less
17
US20170166904A1
Publication/Patent Number: US20170166904A1
Publication date: 2017-06-15
Application number: 15/371,343
Filing date: 2016-12-07
Abstract: The purpose of the invention is to provide means with which it is possible to efficiently select a vector to which a foreign gene has been introduced when a foreign gene is to be introduced by homologous recombination to a vector having multiple sequences homologous with one another. The vector comprises, in succession, a replication origin, a sequence A, a marker gene X, two sequences C and D for introducing a foreign gene by homologous recombination, and a sequence B homologous with sequence A. The two sequences C and D are directly or indirectly adjacent to one another. The vector is used for introducing a foreign gene between the two adjacent sequences C and D. The purpose of the invention is to provide means with which it is possible to efficiently select a vector to which a foreign gene has been introduced when a foreign gene is to be introduced by homologous recombination to a vector having multiple sequences homologous with one ...more ...less
18
US9625464B2
Publication/Patent Number: US9625464B2
Publication date: 2017-04-18
Application number: 13/637,025
Filing date: 2011-03-22
Abstract: A method which can isolate plasma cells and plasmablasts efficiently and with high purity, from mammals and birds, without using a cell surface marker is provided. Further disclosed is a fluorescent probe wherein the staining selectivity for the endoplasmic reticulum of cells is higher than the staining selectivity for cell organelles other than the endoplasmic reticulum. Also disclosed is a method for identifying plasma cells and plasmablasts which includes staining cells derived from lymph node tissue or similar by using this probe, and identifying plasma cells and plasmablasts on the basis of the fluorescence intensity from the stained cells. Also disclosed is a fluorescent probe wherein the staining selectivity for cell nuclei is higher than the staining selectivity for cell organelles other than the cell nuclei. Also disclosed is a method for identifying plasma cells and plasmablasts in lymph node tissue or similar which includes staining cells derived from lymph node tissue or similar by using this probe, and identifying plasma cells and plasmablasts on the basis of the fluorescence intensity from the stained cells. A method which can isolate plasma cells and plasmablasts efficiently and with high purity, from mammals and birds, without using a cell surface marker is provided. Further disclosed is a fluorescent probe wherein the staining selectivity for the endoplasmic reticulum of cells ...more ...less