公开(公告)号：US20070112181A1

公开(公告)日：2007-05-17

申请号：US10582882

申请日：2004-12-21

申请人： Makoto Watanabe ,  Eiichi Matsuo ,  Chikako Toda ,  Osamu Nishimura

发明人： Makoto Watanabe ,  Eiichi Matsuo ,  Chikako Toda ,  Osamu Nishimura

IPC分类号： C07K1/00

CPC分类号： C07K1/24

摘要： The present invention finds a media that can highly selectively retain proteins or peptides to be enriched/separated, as well as provides a method for selectively enriching/separating proteins or peptides using such a media. A method for enrichment/separation of a protein or a peptide, comprising separating a protein or a peptide containing an amino acid residue with a π electron-containing group by using a media with a π electron-containing group. Preferably, the amino acid residue is a tryptophan residue or a tryptophan residue modified with a sulfenyl compound, and the media is a media with phenyl group.

摘要翻译： 本发明找到能够高度选择性保留待富集/分离的蛋白质或多肽的培养基，以及提供使用这种培养基选择性富集/分离蛋白质或肽的方法。 用于富集/分离蛋白质或肽的方法，包括通过使用具有含π电子的基团的培养基来分离含有氨基酸残基的蛋白质或含有含π电子的基团的肽。 优选地，氨基酸残基是用巯基化合物修饰的色氨酸残基或色氨酸残基，并且培养基是具有苯基的培养基。

公开(公告)号：US07354996B2

公开(公告)日：2008-04-08

申请号：US11028322

申请日：2005-01-04

申请人： Eiichi Matsuo ,  Makoto Watanabe ,  Chikako Toda ,  Osamu Nishimura

发明人： Eiichi Matsuo ,  Makoto Watanabe ,  Chikako Toda ,  Osamu Nishimura

IPC分类号： A61K38/00

CPC分类号： G01N33/6842 ,  G01N33/6851

摘要： The present invention provides a method for global quantitative analysis of protein that is effectively applied also for unpurified samples such as biological samples, and achieves better detection sensitivity and quantitativeness than the conventional NBS method. A method for global quantitative analysis of protein comprising: preparing two states of protein samples, a Protein sample I for analysis and a control Protein sample II; solubilizing the Protein sample I and II by urea or guanidine hydrochloride; subjecting the solubilized Protein sample I and II to modification using 2-nitro[13C6]benzenesulfenyl chloride and 2-nitro[12C6]benzenesulfenyl chloride; mixing and desalting the modified Protein sample I and II; resolubilizing by urea or guanidine hydrochloride; reducing and alkylating; subjecting to trypsin digestion in the presence of urea or guanidine hydrochloride; separating the obtained peptide mixture using a media having a phenyl group; and subjecting the enriched modified peptide fragments to mass spectrometry preferably using 3CHCA, 3H4NBA or mixture of 3H4NBA and 4CHCA as a matrix.

摘要翻译： 本发明提供了一种全球定量分析蛋白质的方法，该方法也有效应用于未纯化的样品如生物样品，并且比常规NBS方法获得更好的检测灵敏度和定量性。 一种用于蛋白质全球定量分析的方法，包括：制备蛋白质样品的两种状态，用于分析的蛋白质样品I和对照蛋白质样品II; 通过尿素或盐酸胍溶解蛋白质样品I和II; 将溶解的蛋白质样品I和II用2-硝基[13 C 6]苯硫基氯和2-硝基[12β] 亚苯基氯; 混合和脱盐改性蛋白质样品I和II; 用尿素或盐酸胍重新溶解; 还原和烷基化; 在尿素或盐酸胍存在下进行胰蛋白酶消化; 使用具有苯基的介质分离得到的肽混合物; 并将富集的经修饰的肽片段进行质谱，优选使用3CHCA，3H4NBA或3H4NBA和4CHCA的混合物作为基质。

公开(公告)号：US20060243899A1

公开(公告)日：2006-11-02

申请号：US11028230

申请日：2005-01-04

申请人： Eiichi Matsuo ,  Chikako Toda ,  Makoto Watanabe ,  Osamu Nishimura ,  Noriyuki Ojima

发明人： Eiichi Matsuo ,  Chikako Toda ,  Makoto Watanabe ,  Osamu Nishimura ,  Noriyuki Ojima

IPC分类号： B01D59/44

CPC分类号： H01J49/0418 ,  G01N33/6842 ,  G01N33/6851

摘要： The present invention provides a method which is capable of mainly detecting the objective substance by mass spectrometry and can also be applied effectively to a sample to be measured without conducting enrichment process. A mass spectrometric method comprising: in a mass spectrometry specifically ionizing a specific substance to be measured contained in a mixture sample containing the specific substance and a substance other than the specific substance by using a matrix that is more likely to ionize the specific substance than the substance other than the specific substance, to selectively measure the specific substance from the mixture. Preferably, the specific substance is a peptide modified with 2-nitrobenzenesulfenyl chloride, and the matrix is a nitrobenzene derivative such as hydroxynitrobenzoic acid. The matrix is preferably used in combination with α-cyano-4-hydroxycinnamic acid.

摘要翻译： 本发明提供能够通过质谱法主要检测目标物质的方法，并且还可以在不进行富集过程的情况下有效地应用于要测量的样品。 一种质谱法，其特征在于，在通过使用比所述特定物质更可能离子化的基质更特别地电离含有特定物质的混合物样品和特定物质以外的物质的特定被测物质进行质谱分析 从特定物质以外的物质，选择性地测量混合物中的特定物质。 优选地，特定物质是用2-硝基苯硫酰氯修饰的肽，并且基质是硝基苯衍生物如羟基硝基苯甲酸。 该基质优选与α-氰基-4-羟基肉桂酸组合使用。

公开(公告)号：US20050224710A1

公开(公告)日：2005-10-13

申请号：US11028216

申请日：2005-01-04

申请人： Eiichi Matsuo ,  Makoto Watanabe ,  Noriyuki Ojima ,  Chikako Toda ,  Hiroki Kuyama ,  Osamu Nishimura

发明人： Eiichi Matsuo ,  Makoto Watanabe ,  Noriyuki Ojima ,  Chikako Toda ,  Hiroki Kuyama ,  Osamu Nishimura

IPC分类号： G01N33/68 ,  H01J49/16

CPC分类号： H01J49/164 ,  G01N33/6848

摘要： The present invention provides a method capable of efficiently ionizing hydrophobic peptides in MALDI-IT, MALDI-IT-TOF, and MALDI-FTICR mass spectrometers. A method of measuring a peptide with a mass spectrometer having a MALDI (Matrix Assisted Laser Desorption/Ionization) ion source, using α-cyano-3-hydroxycinnamic acid or 3-hydroxy-4-nitrobenzoic acid as a matrix. Preferably, a peptide derivatized with 2-nitrobenzenesulfenyl chloride is measured with a MALDI-IT, MALDI-IT-TOF, or MALDI-FTICR mass spectrometer. When 3-hydroxy-4-nitrobenzoic acid is used as a matrix, the matrix is preferably used as a mixed matrix in which α-cyano-4-hydroxycinnamic acid is combined.

摘要翻译： 本发明提供了能够有效地离子化MALDI-IT，MALDI-IT-TOF和MALDI-FTICR质谱仪中的疏水性肽的方法。 使用α-氰基-3-羟基肉桂酸或3-羟基-4-硝基苯甲酸作为基质的具有MALDI（Matrix Assisted Laser Desorption / Ionization）离子源的质谱仪测量肽的方法。 优选地，用MALDI-IT，MALDI-IT-TOF或MALDI-FTICR质谱仪测量用2-硝基苯硫基氯衍生的肽。 当使用3-羟基-4-硝基苯甲酸作为基质时，优选将基质用作混合基质，其中组合了α-氰基-4-羟基肉桂酸。

公开(公告)号：US20060051831A1

公开(公告)日：2006-03-09

申请号：US11028322

申请日：2005-01-04

申请人： Eiichi Matsuo ,  Makoto Watanabe ,  Chikako Toda ,  Osamu Nishimura

发明人： Eiichi Matsuo ,  Makoto Watanabe ,  Chikako Toda ,  Osamu Nishimura

IPC分类号： C12Q1/37

CPC分类号： G01N33/6842 ,  G01N33/6851

摘要： The present invention provides a method for global quantitative analysis of protein that is effectively applied also for unpurified samples such as biological samples, and achieves better detection sensitivity and quantitativeness than the conventional NBS method. A method for global quantitative analysis of protein comprising: preparing two states of protein samples, a Protein sample I for analysis and a control Protein sample II; solubilizing the Protein sample I and II by urea or guanidine hydrochloride; subjecting the solubilized Protein sample I and II to modification using 2-nitro[13C6]benzenesulfenyl chloride and 2-nitro[12C6]benzenesulfenyl chloride; mixing and desalting the modified Protein sample I and II; resolubilizing by urea or guanidine hydrochloride; reducing and alkylating; subjecting to trypsin digestion in the presence of urea or guanidine hydrochloride; separating the obtained peptide mixture using a media having a phenyl group; and subjecting the enriched modified peptide fragments to mass spectrometry preferably using 3CHCA, 3H4NBA or mixture of 3H4NBA and 4CHCA as a matrix.

摘要翻译： 本发明提供了一种全球定量分析蛋白质的方法，该方法也有效应用于未纯化的样品如生物样品，并且比常规NBS方法获得更好的检测灵敏度和定量性。 一种用于蛋白质全球定量分析的方法，包括：制备蛋白质样品的两种状态，用于分析的蛋白质样品I和对照蛋白质样品II; 通过尿素或盐酸胍溶解蛋白质样品I和II; 将溶解的蛋白质样品I和II用2-硝基[13 C 6]苯硫基氯和2-硝基[12β] 亚苯基氯; 混合和脱盐改性蛋白质样品I和II; 用尿素或盐酸胍重新溶解; 还原和烷基化; 在尿素或盐酸胍存在下进行胰蛋白酶消化; 使用具有苯基的介质分离得到的肽混合物; 并将富集的经修饰的肽片段进行质谱，优选使用3CHCA，3H4NBA或3H4NBA和4CHCA的混合物作为基质。

公开(公告)号：US20130065258A1

公开(公告)日：2013-03-14

申请号：US13699592

申请日：2011-01-28

申请人： Makoto Watanabe ,  Ei-ichi Matsuo ,  Naoki Kaneko ,  Toshiya Matsubara ,  Osamu Nishimura ,  Masaki Mori ,  Ichiro Takemasa

发明人： Makoto Watanabe ,  Ei-ichi Matsuo ,  Naoki Kaneko ,  Toshiya Matsubara ,  Osamu Nishimura ,  Masaki Mori ,  Ichiro Takemasa

IPC分类号： G01N33/574 ,  C07K14/47 ,  G01N21/64

CPC分类号： G01N33/57419 ,  G01N2333/4724

摘要： The present invention provides a tumor screening marker that can be actually used in clinical practice to detect colorectal cancer, and a tumor progression marker that can complement CEA or CA19-9. Galectin-1 used as a tumor screening marker or a tumor progression marker for colorectal cancer. Galectin-3 used as a tumor screening marker. Galectin-4 used as a tumor progression marker, a tumor screening marker, or a prognostic prediction marker for colorectal cancer. A method of analyzing the galectin concentration in a collected blood sample using the galectin. A colorectal cancer marker detection kit comprising a detection antibody selected from the group consisting of a fluorescently labeled galectin-1 antibody, a fluorescently labeled galectin-3 antibody, and a fluorescently labeled galectin-4 antibody.

摘要翻译： 本发明提供可以实际用于临床实践中检测结肠直肠癌的肿瘤筛选标记，以及可补充CEA或CA19-9的肿瘤进展标记。 Galectin-1用作肿瘤筛选标记物或肿瘤进展标志物用于结肠直肠癌。 Galectin-3用作肿瘤筛选标记。 用作肿瘤进展标记物的Galectin-4，肿瘤筛选标记或结肠直肠癌的预后预测标记物。 使用半乳凝素分析收集的血液样品中的半乳凝素浓度的方法。 一种包含选自荧光标记的半乳凝素-1抗体，荧光标记的半乳凝素-3抗体和荧光标记的半乳凝素-4抗体的检测抗体的结肠直肠癌标志物检测试剂盒。

公开(公告)号：US20120149026A1

公开(公告)日：2012-06-14

申请号：US13370840

申请日：2012-02-10

申请人： Toshiya Matsubara ,  Osamu Nishimura ,  Susumu Iwasa ,  Makoto Watanabe

发明人： Toshiya Matsubara ,  Osamu Nishimura ,  Susumu Iwasa ,  Makoto Watanabe

IPC分类号： C12Q1/68 ,  G01N33/53 ,  C07K16/18 ,  C07H21/04 ,  C12Q1/37 ,  G01N33/566

CPC分类号： C07K14/52 ,  C07K14/475 ,  C12Q1/6883 ,  C12Q2600/136 ,  C12Q2600/158 ,  G01N33/68 ,  G01N2500/04 ,  G01N2500/10 ,  G01N2800/042

摘要： It is provided an insulin resistance marker, a method of evaluating insulin resistance, a method of screening a substance that improves insulin resistance, and a pharmaceutical composition for improving insulin resistance. An insulin resistance marker including a polypeptide comprising at least any 15 continuous amino acids in the specific amino acid sequence (sequence of a proepithelin protein). An insulin resistance marker including a polynucleotide selected from the group consisting of a polynucleotide comprising at least any 45 continuous bases in the base sequence encoding the above specific amino acid sequence, and a polynucleotide that is complementary to the polynucleotide.

摘要翻译： 本发明提供胰岛素抵抗标记，胰岛素抵抗评价方法，筛选改善胰岛素抵抗的物质的方法以及改善胰岛素抵抗的药物组合物。 包括在特定氨基酸序列（前列腺素蛋白的序列）中包含至少15个连续氨基酸的多肽的胰岛素抵抗标记。 包括选自包含编码上述特定氨基酸序列的碱基序列中至少任何45个连续碱基的多核苷酸的多核苷酸的多核苷酸和与该多核苷酸互补的多核苷酸的胰岛素抵抗标记。

公开(公告)号：US20090191575A1

公开(公告)日：2009-07-30

申请号：US12303363

申请日：2007-06-05

申请人： Makoto Watanabe ,  Osamu Nishimura ,  Toshiya Matsubara ,  Ichiro Takemasa ,  Morito Monden ,  Katsuya Nagai ,  Nariaki Matsuura

发明人： Makoto Watanabe ,  Osamu Nishimura ,  Toshiya Matsubara ,  Ichiro Takemasa ,  Morito Monden ,  Katsuya Nagai ,  Nariaki Matsuura

IPC分类号： G01N33/574 ,  C12N9/12 ,  C12N9/24 ,  C07K14/00 ,  C12N9/10 ,  C12N9/14 ,  C12N9/50 ,  C12N9/02 ,  G01N33/573

CPC分类号： G01N33/57419

摘要： The present invention provides a tumor marker and a method capable of identifying the morbidity of colon cancer. A tumor marker including a protein identified from a colon cancer tissue. A method for identify the morbidity of colon cancer using the tumor marker. The method includes: measuring the level of the protein in a sample derived from a person of interest who should be examined to identify the morbidity of colon cancer; and comparing the measured level to the normal level of the protein, wherein a higher or lower measured level than the normal level is used as one indicator indicating that there is a high possibility that the person of interest has colon cancer.

摘要翻译： 本发明提供了能够鉴定结肠癌发病率的肿瘤标志物和方法。 包括从结肠癌组织鉴定的蛋白质的肿瘤标志物。 使用肿瘤标志物鉴定结肠癌发病率的方法。 该方法包括：测量衍生自感兴趣的人的样品中蛋白质的水平，其应当被检查以鉴定结肠癌的发病率; 并且将测量的水平与蛋白质的正常水平进行比较，其中使用比正常水平更高或更低的测量水平作为一个指标，指示感兴趣的人具有结肠癌的可能性很高。

公开(公告)号：US09079972B2

公开(公告)日：2015-07-14

申请号：US13370840

申请日：2012-02-10

申请人： Toshiya Matsubara ,  Osamu Nishimura ,  Susumu Iwasa ,  Makoto Watanabe

发明人： Toshiya Matsubara ,  Osamu Nishimura ,  Susumu Iwasa ,  Makoto Watanabe

IPC分类号： G01N33/53 ,  C07K14/52 ,  C07K14/475 ,  C12Q1/68 ,  G01N33/68

CPC分类号： C07K14/52 ,  C07K14/475 ,  C12Q1/6883 ,  C12Q2600/136 ,  C12Q2600/158 ,  G01N33/68 ,  G01N2500/04 ,  G01N2500/10 ,  G01N2800/042

摘要： An insulin resistance marker, a method of evaluating insulin resistance, a method of screening a substance that improves insulin resistance, and a pharmaceutical composition for improving insulin resistance are provided. The insulin resistance marker includes a polypeptide comprising at least any 15 continuous amino acids in the specific amino acid sequence of SEQ ID NO: 1 (sequence of a proepithelin protein). The insulin resistance marker includes a polynucleotide selected from the group consisting of (1) a polynucleotide comprising at least any 45 continuous bases in the base sequence of SEQ ID NO:2 encoding the above specific amino acid sequence, and (2) a polynucleotide that is complementary to the polynucleotide of (1).

摘要翻译： 提供胰岛素抵抗标记，评价胰岛素抵抗的方法，筛选改善胰岛素抵抗的物质的方法和用于改善胰岛素抵抗的药物组合物。 胰岛素抵抗标记物包括在SEQ ID NO：1的特定氨基酸序列（前列腺素蛋白质序列）中包含至少15个连续氨基酸的多肽。 胰岛素抵抗标记物包括选自（1）包含编码上述特定氨基酸序列的SEQ ID NO：2的碱基序列中至少任何45个连续碱基的多核苷酸的多核苷酸，和（2）多核苷酸， 与（1）的多核苷酸互补。

公开(公告)号：US20130071864A1

公开(公告)日：2013-03-21

申请号：US13699608

申请日：2011-01-28

申请人： Makoto Watanabe ,  Ei-Ichi Matsuo ,  Naoki Kaneko ,  Toshiya Matsubara ,  Masaki Mori ,  Ichiro Takemasa ,  Osamu Nishimura

发明人： Makoto Watanabe ,  Ei-Ichi Matsuo ,  Naoki Kaneko ,  Toshiya Matsubara ,  Masaki Mori ,  Ichiro Takemasa ,  Osamu Nishimura

IPC分类号： G01N33/574 ,  C07K14/435

CPC分类号： G01N33/57419 ,  G01N2333/78

摘要： The present invention provides a tumor screening marker that can be actually used in clinical practice to detect colorectal cancer, and a tumor progression marker that can complement CEA or CA19-9. Vitronectin for use as a tumor progression marker, a tumor screening marker or a prognostic prediction marker for colorectal cancer. A method of analyzing a vitronectin concentration in a collected blood sample. In the method, a measured value of vitronectin and a reference value of vitronectin are compared. Vitronectin is preferably used in combination with existing marker for colorectal cancer such as carcinoembryonic antigen and CA19-9.

摘要翻译： 本发明提供可以实际用于临床实践中检测结肠直肠癌的肿瘤筛选标记，以及可补充CEA或CA19-9的肿瘤进展标记。 用作肿瘤进展标记的Vitronectin，肿瘤筛选标记或结肠直肠癌的预后预测标记。 分析收集血液样品中玻连蛋白浓度的方法。 在该方法中，比较玻连蛋白的测定值和玻连蛋白的参考值。 玻连蛋白优选与现有的结肠直肠癌标记物如癌胚抗原和CA19-9组合使用。