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：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：US20060046303A1

公开(公告)日：2006-03-02

申请号：US10531288

申请日：2003-04-03

申请人： Hiroki Kuyama ,  Chikako Toda ,  Osamu Nishimura

发明人： Hiroki Kuyama ,  Chikako Toda ,  Osamu Nishimura

IPC分类号： G01N1/00

CPC分类号： G01N33/6848 ,  C07K1/113 ,  C07K7/06 ,  C07K7/08 ,  G01N33/6842 ,  G01N33/6851 ,  Y10T436/25125

摘要： A method for eliminating a phosphate group of a peptide, the method comprising the use of a reagent containing at least one selected from the group consisting of hydrogen fluoride, hydrofluoric acid, and a hydrogen fluoride-containing compound, and a method for peptide analysis that uses such a method. As the hydrogen fluoride-containing compound, hydrogen fluoride-pyridine is preferably used. The elimination of the phosphate group from a peptide may be carried out so that the total amount of the hydrogen fluoride, hydrogen fluoride in the hydrofluoric acid, and hydrogen fluoride in the hydrogen fluoride-containing compound contain in the reagent is 10 to 100 wt % with respect to the reagent, and the temperature for the elimination reaction is −10 to 50° C.

摘要翻译： 一种消除肽的磷酸基团的方法，该方法包括使用含有选自氟化氢，氢氟酸和含氟化氢的化合物中的至少一种的试剂，以及肽分析方法， 使用这种方法。 作为含氟化氢的化合物，优选使用氟化氢 - 吡啶。 可以进行磷酸基从肽的去除，使得在试剂中含有的氟化氢，氢氟酸中的氟化氢和含氟化氢的化合物中的氟化氢的总量为10〜100重量％ 相对于试剂，消除反应的温度为-10〜50℃。

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。