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1. 发明申请

US20150204870A1 UNDIFFERENTIATED CELL DETECTION METHOD AND COMPLEX CARBOHYDRATE DETECTION METHOD 有权
标题翻译：不稳定细胞检测方法和复合碳水化合物检测方法
公开(公告)号：US20150204870A1
公开(公告)日：2015-07-23
申请号：US14355413
申请日：2012-10-31
申请人： NATIONAL INSTITUTE OF ADVANCED INDUSTRIAL SCIENCE AND TECHNOLOGY , WAKO PURE CHEMICAL INDUSTRIES, LTD.
发明人： Hiroaki Tateno , Jun Hirabayashi , Yuzuru Ito , Yasuko Onuma , Makoto Asashima , Atsushi Kuno , Masaki Warashina , Masakazu Fukuda
IPC分类号： G01N33/569 , C07K14/195
CPC分类号： G01N33/56966 , C07K14/195 , G01N2333/4724 , G01N2400/00
摘要： An object of the present invention is to provide a method for evaluating a differentiation status of cells using a culture supernatant of stem cells. Provided is “undifferentiation sugar chain marker” composed of the sugar chain structure “Fucα1-2Galβ1-3GlcNAc” or “Fucα1-2Galβ1-3GalNAc” and capable of sensitively determining the undifferentiated state of stem cells using a culture supernatant of stem cells. Also found is BC2LCN lectin or a modified product thereof capable of sensitively recognizing the “undifferentiation sugar chain marker” as excellent “probe for detecting the undifferentiation sugar chain marker” capable of determining the undifferentiation status of cells using a culture supernatant.
摘要翻译：本发明的目的是提供一种使用干细胞的培养物上清液评估细胞分化状态的方法。提供由糖链结构“Fucα1-2Gal＆bgr; 1-3GlcNAc”或“Fucα1-2Gal＆bgr; 1-3GalNAc”组成的“未分化糖链标记”，其能够使用茎的培养物上清液敏感地测定干细胞的未分化状态细胞。还发现BC2LCN凝集素或其能够敏感识别“未分化糖链标记”的优化“能够用培养上清液测定细胞未分化状态的”未分化糖链标志物的探针“的修饰产物。

2. 发明授权

US09279809B2 Cell differentiation assay method, cell isolation method, method for producing induced pluripotent stem cells, and method for producing differentiated cells 有权
标题翻译：细胞分化试验方法，细胞分离方法，诱导多能干细胞的制备方法和分化细胞的制备方法
公开(公告)号：US09279809B2
公开(公告)日：2016-03-08
申请号：US14381116
申请日：2013-02-27
申请人： National Institute of Advanced Industrial Science and Technology , Wako Pure Chemical Industries, Ltd.
发明人： Hiroaki Tateno , Jun Hirabayashi , Makoto Asashima , Yuzuru Ito , Yasuko Onuma , Masaki Warashina , Masakazu Fukuda
IPC分类号： G01N33/569
CPC分类号： G01N33/56966
摘要： Provided are a method for accurately evaluating the differentiation status of stem cells by selectively staining only stem cells in an undifferentiated state, and a method for positively isolating only stem cells in an undifferentiated state. Specifically provided is a method for determining differentiation of a cell comprising a step of contacting a test cell with a probe comprising protein (A) or (B) below and a step of detecting the presence of binding of the probe to the test cell. The method for determining differentiation of a cell is capable of detecting the presence or absence of an undifferentiated stem cell in test cells by using a probe that specifically reacts with undifferentiated stem cells and detecting the presence of bonding to the test cell. (A) A protein comprising an amino acid sequence shown in SEQ ID NO: 1 and recognizing a sugar chain structure of “Fucα1-2Galβ1-3GlcNAc” and/or “Fucα1-2Galβ1-3GalNAc;” and (B) a protein comprising an amino acid sequence showing 80% or more similarity to the amino acid sequence shown in SEQ ID NO: 1 and recognizing a sugar chain structure of “Fucα1-2Galβ1-3GlcNAc” and/or “Fucα1-2Galβ1-3GalNAc.”
摘要翻译：提供了通过仅选择性染色未分化状态的干细胞来精确评估干细胞的分化状态的方法，以及仅将未分化状态的干细胞仅阳性分离的方法。具体提供了一种用于确定细胞分化的方法，包括使测试细胞与包含以下蛋白质（A）或（B）的探针接触的步骤和检测探针与测试细胞结合的步骤的步骤。用于确定细胞分化的方法能够通过使用与未分化干细胞特异性反应并检测与测试细胞结合的存在的探针来检测测试细胞中是否存在未分化干细胞。（A）包含SEQ ID NO：1所示的氨基酸序列并识别“Fucα1-2Gal”和“3GlcNAc”和/或“Fucα1-2Gal”和“1-3GalNAc”的糖链结构的蛋白质和（B）包含与SEQ ID NO：1所示的氨基酸序列具有80％以上相似性的氨基酸序列的蛋白质，并且识别“Fucα1-2Gal”和“3GlcNAc”的糖链结构和/或“Fucα1-2Gal” -3GalNAc“。

3. 发明申请

US20150111218A1 CELL DIFFERENTIATION ASSAY METHOD, CELL ISOLATION METHOD, METHOD FOR PRODUCING INDUCED PLURIPOTENT STEM CELLS, AND METHOD FOR PRODUCING DIFFERENTIATED CELLS 有权
标题翻译：细胞分化测定方法，细胞分离方法，产生诱导型细胞干细胞的方法和生产差异细胞的方法
公开(公告)号：US20150111218A1
公开(公告)日：2015-04-23
申请号：US14381116
申请日：2013-02-27
申请人： National Institute of Advanced Industrial Science and Technology , Wako Pure Chemical Industries, Ltd.
发明人： Hiroaki Tateno , Jun Hirabayashi , Makoto Asashima , Yuzuru Ito , Yasuko Onuma , Masaki Warashina , Masakazu Fukuda
IPC分类号： G01N33/569
CPC分类号： G01N33/56966
摘要： Provided are a method for accurately evaluating the differentiation status of stem cells by selectively staining only stem cells in an undifferentiated state, and a method for positively isolating only stem cells in an undifferentiated state. Specifically provided is a method for determining differentiation of a cell comprising a step of contacting a test cell with a probe comprising protein (A) or (B) below and a step of detecting the presence of binding of the probe to the test cell. The method for determining differentiation of a cell is capable of detecting the presence or absence of an undifferentiated stem cell in test cells by using a probe that specifically reacts with undifferentiated stem cells and detecting the presence of bonding to the test cell. (A) A protein comprising an amino acid sequence shown in SEQ ID NO: 1 and recognizing a sugar chain structure of “Fucα1-2Galβ1-3GlcNAc” and/or “Fucα1-2Galβ1-3GalNAc;” and (B) a protein comprising an amino acid sequence showing 80% or more similarity to the amino acid sequence shown in SEQ ID NO: 1 and recognizing a sugar chain structure of “Fucα1-2Galβ1-3GlcNAc” and/or “Fucα1-2Galβ1-3GalNAc.”
摘要翻译：提供了通过仅选择性染色未分化状态的干细胞来精确评估干细胞的分化状态的方法，以及仅将未分化状态的干细胞仅阳性分离的方法。具体提供了一种用于确定细胞分化的方法，包括使测试细胞与包含以下蛋白质（A）或（B）的探针接触的步骤和检测探针与测试细胞结合的步骤的步骤。用于确定细胞分化的方法能够通过使用与未分化干细胞特异性反应并检测与测试细胞结合的存在的探针来检测测试细胞中是否存在未分化干细胞。（A）包含SEQ ID NO：1所示的氨基酸序列并识别“Fucα1-2Gal”和“3GlcNAc”和/或“Fucα1-2Gal”和“1-3GalNAc”的糖链结构的蛋白质和（B）包含与SEQ ID NO：1所示的氨基酸序列具有80％以上相似性的氨基酸序列的蛋白质，并且识别“Fucα1-2Gal”和“3GlcNAc”的糖链结构和/或“Fucα1-2Gal” -3GalNAc“。

4. 发明授权

US09500650B2 Undifferentiated cell detection method and complex carbohydrate detection method 有权
标题翻译：未分化细胞检测方法和复杂碳水化合物检测方法
公开(公告)号：US09500650B2
公开(公告)日：2016-11-22
申请号：US14355413
申请日：2012-10-31
申请人： National Institute of Advanced Industrial Science and Technology , Wako Pure Chemical Industries, Ltd.
发明人： Hiroaki Tateno , Jun Hirabayashi , Yuzuru Ito , Yasuko Onuma , Makoto Asashima , Atsushi Kuno , Masaki Warashina , Masakazu Fukuda
IPC分类号： G01N33/569 , C07K14/195
CPC分类号： G01N33/56966 , C07K14/195 , G01N2333/4724 , G01N2400/00
摘要： An object of the present invention is to provide a method for evaluating a differentiation status of cells using a culture supernatant of stem cells. Provided is “undifferentiation sugar chain marker” composed of the sugar chain structure “Fucα1-2Galβ1-3GlcNAc” or “Fucα1-2Galβ1-3GalNAc” and capable of sensitively determining the undifferentiated state of stem cells using a culture supernatant of stem cells. Also found is BC2LCN lectin or a modified product thereof capable of sensitively recognizing the “undifferentiation sugar chain marker” as excellent “probe for detecting the undifferentiation sugar chain marker” capable of determining the undifferentiation status of cells using a culture supernatant.
摘要翻译：本发明的目的是提供一种使用干细胞的培养物上清液评估细胞分化状态的方法。提供由糖链结构“Fucα1-2Galβ1-3GlcNAc”或“Fucα1-2Galβ1-3GalNAc”组成的能够使用干细胞的培养上清液敏感地测定干细胞的未分化状态的“未分化糖链标记”。还发现BC2LCN凝集素或其能够敏感识别“未分化糖链标记”的优化“能够用培养上清液测定细胞未分化状态的”未分化糖链标志物的探针“的修饰产物。

5. 发明申请

US20150329602A1 MODIFIED LECTIN DERIVED FROM WISTERIA FLORIBUNDA 有权
标题翻译：来自WISTERIA FLORIBUNDA的改良的LECTIN
公开(公告)号：US20150329602A1
公开(公告)日：2015-11-19
申请号：US14654223
申请日：2013-12-18
申请人： NATIONAL INSTITUTE OF ADVANCED INDUSTRIAL SCIENCE AND TECHNOLOGY
发明人： Takashi Sato , Yasunori Chiba , Hiroaki Tateno , Hiroyuki Kaji , Masanori Goto , Hisashi Narimatsu
IPC分类号： C07K14/42 , G01N33/53
CPC分类号： C07K14/42 , G01N33/5308 , G01N2333/42 , G01N2400/00
摘要： [Problem] The purpose of the present invention is to stably supply high-quality and highly uniform Wisteria floribunda agglutinin (WFA) that recognizes biologically important sugar-chain markers, to elucidate the sugar-chain recognition activity in detail, and to furthermore increase the specificity of the sugar-chain recognition activity. [Solution] The present invention involves the development of a technique for cloning genes for coding Wisteria floribunda agglutinin (WFA) and producing recombinant WFA having the same sugar-chain recognition activity as natural WFA from transformed bacteria. Natural WFA is reduced to thereby manufacture a reduced WFA monomer for specifically recognizing terminal GalNAc residue. A recombinant monomer WFA for recognizing LDN (GalNAcβ1, 4GlcNAc) sugar chain, which is important as a diagnostic marker among sugar chains having a terminal GalNAc residue, is manufactured by introducing cysteine mutation to recombinant WFA or by C-terminal-side amino acid deletion.
摘要翻译： [问题]本发明的目的是稳定地提供识别生物重要的糖链标记物的高品质和高度均匀的紫藤凝集素（WFA），以详细阐明糖链识别活性，并进一步增加糖链识别活性的特异性。 [解决方案]本发明涉及开发克隆用于编码Wisteria floribunda凝集素（WFA）的基因的克隆技术，并且生产与来自转化细菌的天然WFA具有相同糖链识别活性的重组WFA。减少天然WFA，从而制造用于特异性识别末端GalNAc残基的还原的WFA单体。通过将半胱氨酸突变引入重组WFA或C末端侧氨基酸来制备重要单体WFA，其用于识别LDN（GalNAc＆bgr; 1,4GlcNAc）糖链，其作为具有末端GalNAc残基的糖链中的诊断标记是重要的酸缺失。

6. 发明授权

US11078254B2 Method for immobilizing lectin 有权
公开(公告)号：US11078254B2
公开(公告)日：2021-08-03
申请号：US16604565
申请日：2018-04-11
申请人： National Institute of Advanced Industrial Science and Technology , Nissan Chemical Corporation , National University Corporation Kyoto Institute of Technology
发明人： Hiroaki Tateno , Junko Katayama , Kazutaka Matoba , Yoichi Kumada
IPC分类号： C07K7/06 , C07K14/195 , C07K17/08 , C07K17/14 , C07K19/00 , G01N33/545 , G01N33/551 , G01N33/566 , C07K14/705 , C07K17/00 , C12N15/62 , G01N33/543
摘要： Provided is a highly sensitive and less expensive lectin-immobilized base material (for example, a lectin plate), such as lectin-immobilized base material having stable qualities and being able to be sufficiently washed after a target sugar chain-containing antigen binds thereto. Further provided is a method for immobilizing lectin to a base material therefor. Particularly provided are: a method whereby a lectin-peptide fusion, in which a peptide capable of adsorbing to a base material surface such as a polystyrene (PS) tag is fused with the N-terminal side or C-terminal side of lectin capable of recognizing a target sugar chain, is immobilized on the peptide side to a base material; and a lectin-immobilized base material produced by this method. By using the lectin-immobilized base material, a target sugar chain-containing antigen can be highly sensitively and evenly measured and, moreover, target sugar chain-containing cells, etc. can be separated (concentrated and harvested).

7. 发明授权

US10539553B2 Method and kit for detecting stem cell 有权
公开(公告)号：US10539553B2
公开(公告)日：2020-01-21
申请号：US15548921
申请日：2015-12-21
申请人： NATIONAL INSTITUTE OF ADVANCED INDUSTRIAL SCIENCE AND TECHNOLOGY , FUJIFILM WAKO PURE CHEMICAL CORPORATION
发明人： Hiroaki Tateno , Masaki Warashina , Masakazu Fukuda , Kazunari Hirayasu
IPC分类号： G01N33/53 , G01N33/50 , C07H5/04 , G01N33/574
摘要： Provided is a method for detecting a stem cell based on an undifferentiated sugar chain marker having a specific sugar chain structure, wherein the stem cell is detected by detecting podocalyxin contained in a culture supernatant or a lysate of cells by a “lectin-antibody sandwich method” using a combination of a lectin and an antibody and having high sensitivity, the method including steps of: contacting the culture supernatant or the lysate, a lectin capable of binding to a sugar chain represented by (Formula 1) or (Formula 2), and an antibody capable of binding to keratan sulfate to form a complex composed of the lectin, podocalyxin and the antibody; and detecting the complex. wherein R1 represents an OH group or any sugar chain and R2 represents an OH group or any sugar chain, protein, lipid, or another molecule. wherein R1 represents an OH group or any sugar chain and R2 represents an OH group or any sugar chain, protein, lipid, or another molecule.

8. 发明授权

US10358466B2 Modified lectin derived from Wisteria floribunda 有权
公开(公告)号：US10358466B2
公开(公告)日：2019-07-23
申请号：US15706144
申请日：2017-09-15
申请人： NATIONAL INSTITUTE OF ADVANCED INDUSTRIAL SCIENCE AND TECHNOLOGY
发明人： Takashi Sato , Yasunori Chiba , Hiroaki Tateno , Hiroyuki Kaji , Masanori Goto , Hisashi Narimatsu
IPC分类号： C07K14/42 , G01N33/53
摘要： A Wisteria floribunda monomeric lectin polypeptide is provided. The Wisteria floribunda monomeric lectin polypeptide includes any one of the amino acid sequences selected from the group consisting of: (1) the amino acid sequence represented by SEQ ID NO: 2; (2) the amino acid sequence defined in (1) above, except that one to 20 amino acids at positions other than Cys272 position is/are deleted, substituted, inserted, or added; and (3) the amino acid sequence defined in (1) or (2) above, further having an N-terminus deletion of one to 30 amino acids, in which Cys272 is alkylated, and the polypeptide is capable of specifically binding to a GalNAc terminal sugar chain.

9. 发明申请

US20180134756A1 MODIFIED LECTIN DERIVED FROM WISTERIA FLORIBUNDA 审中-公开
公开(公告)号：US20180134756A1
公开(公告)日：2018-05-17
申请号：US15706144
申请日：2017-09-15
申请人： NATIONAL INSTITUTE OF ADVANCED INDUSTRIAL SCIENCE AND TECHNOLOGY
发明人： Takashi Sato , Yasunori Chiba , Hiroaki Tateno , Hiroyuki Kaji , Masanori Goto , Hisashi Narimatsu
IPC分类号： C07K14/42 , G01N33/53
CPC分类号： C07K14/42 , G01N33/5308 , G01N2333/42 , G01N2400/00
摘要： A Wisteria floribunda monomeric lectin polypeptide is provided. The Wisteria floribunda monomeric lectin polypeptide includes any one of the amino acid sequences selected from the group consisting of: (1) the amino acid sequence represented by SEQ ID NO: 2; (2) the amino acid sequence defined in (1) above, except that one to 20 amino acids at positions other than Cys272 position is/are deleted, substituted, inserted, or added; and (3) the amino acid sequence defined in (1) or (2) above, further having an N-terminus deletion of one to 30 amino acids, in which Cys272 is alkylated, and the polypeptide is capable of specifically binding to a GalNAc terminal sugar chain.

10. 发明授权

US09914908B2 Method of introducing a toxin or dye into mammalian pluripotent cells using rBC2LCN 有权
公开(公告)号：US09914908B2
公开(公告)日：2018-03-13
申请号：US14767880
申请日：2014-02-13
申请人： National Institute of Advanced Industrial Science and Technology
发明人： Hiroaki Tateno , Yuzuru Ito , Yasuko Onuma , Jun Hirabayashi , Makoto Asashima
IPC分类号： C12N5/00 , C07K14/195 , C12N9/10
CPC分类号： C12N5/0081 , C07K14/195 , C07K2319/33 , C07K2319/55 , C12N9/1077 , C12Y204/02036
摘要： An object of the present invention is to provide a method for introducing a target substance into undifferentiated cells, and a carrier therefor, whereby the target substance can be specifically introduced into the undifferentiated cells by contacting the undifferentiated cells with an rBC2LCN-target substance fusion product in which rBC2LCN lectin is fused to the target substance. Particularly, an rBC2LCN-toxin fusion product in which rBC2LCN lectin is fused to a toxin functioning in cells or its domain having the ability to kill cells functions as an agent for eliminating undifferentiated stem cells and can be administered into a medium after inducing the differentiation of the stem cells to reliably kill only the stem cells in an undifferentiated state.

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