公开(公告)号：US07892846B2

公开(公告)日：2011-02-22

申请号：US12269336

申请日：2008-11-12

申请人： Hiroki Kuyama ,  Kazuhiro Sonomura ,  Osamu Nishimura

发明人： Hiroki Kuyama ,  Kazuhiro Sonomura ,  Osamu Nishimura

IPC分类号： G01N33/00

CPC分类号： G01N33/6848 ,  Y10T436/16 ,  Y10T436/24

摘要： According to the present invention, a matrix reagent prepared by adding methylenediphosphonic acid (MDPNA) having two phosphonic acid groups as an additive to a matrix of 2,5-dihydroxybenzoic acid (DHBA) is used for the preparation of a sample. By using the sample according to the present invention, it is possible to achieve a higher peak intensity of phosphopeptide as compared with a sample in which DHBA is solely used as a matrix without using an additive, or a sample in which phosphoric acid (PA) is used as an additive in addition to DHBA. Further, use of the sample according to the present invention enables the detection of peptides that cannot be detected in a case of the phosphoric acid-added sample.

摘要翻译： 根据本发明，通过将具有两个膦酸基的亚甲基二膦酸（MDPNA）作为添加剂添加到2,5-二羟基苯甲酸（DHBA）的基质中而制备的基质试剂用于制备样品。 通过使用根据本发明的样品，与仅使用DHBA作为基质而不使用添加剂的样品相比，可以实现更高的磷酸肽峰值强度，或者其中磷酸（PA） 除了DHBA之外还用作添加剂。 此外，根据本发明的样品的使用能够检测在添加磷酸的样品的情况下不能检测到的肽。

公开(公告)号：US20090137052A1

公开(公告)日：2009-05-28

申请号：US12269336

申请日：2008-11-12

申请人： Hiroki KUYAMA ,  Kazuhiro Sonomura ,  Osamu Nishimura

发明人： Hiroki KUYAMA ,  Kazuhiro Sonomura ,  Osamu Nishimura

IPC分类号： G01N33/50

CPC分类号： G01N33/6848 ,  Y10T436/16 ,  Y10T436/24

摘要： According to the present invention, a matrix reagent prepared by adding methylenediphosphonic acid (MDPNA) having two phosphonic acid groups as an additive to a matrix of 2,5-dihydroxybenzoic acid (DHBA) is used for the preparation of a sample. By using the sample according to the present invention, it is possible to achieve a higher peak intensity of phosphopeptide as compared with a sample in which DHBA is solely used as a matrix without using an additive, or a sample in which phosphoric acid (PA) is used as an additive in addition to DHBA. Further, use of the sample according to the present invention enables the detection of peptides that cannot be detected in a case of the phosphoric acid-added sample.

摘要翻译： 根据本发明，通过将具有两个膦酸基的亚甲基二膦酸（MDPNA）作为添加剂添加到2,5-二羟基苯甲酸（DHBA）的基质中而制备的基质试剂用于制备样品。 通过使用根据本发明的样品，与仅使用DHBA作为基质而不使用添加剂的样品相比，可以实现更高的磷酸肽峰值强度，或者其中磷酸（PA） 除了DHBA之外还用作添加剂。 此外，根据本发明的样品的使用能够检测在添加磷酸的样品的情况下不能检测到的肽。

公开(公告)号：US20090142851A1

公开(公告)日：2009-06-04

申请号：US12325115

申请日：2008-11-28

申请人： Keisuke Shima ,  Minoru Yamaguchi ,  Hiroki Kuyama ,  Eiji Ando ,  Osamu Nishimura ,  Susumu Tsunasawa ,  Kazuhiro Sonomura

发明人： Keisuke Shima ,  Minoru Yamaguchi ,  Hiroki Kuyama ,  Eiji Ando ,  Osamu Nishimura ,  Susumu Tsunasawa ,  Kazuhiro Sonomura

IPC分类号： G01N33/68 ,  C07K1/14 ,  C12P21/06

CPC分类号： C07K1/1072 ,  C07K1/128 ,  G01N33/6821

摘要： The present invention provides a method for specifically recovering a C-terminal peptide fragment, and a method for easily determining the sequence of a C-terminal peptide fragment, which is difficult to be determined by a conventional method, with the use of a mass spectrometer, in particular a method capable of de novo sequencing of a C-terminal peptide fragment. A method for selectively recovering a C-terminal peptide of a protein, comprising the steps of: in a cleavage product of a protein containing a C-terminal peptide fragment (A) having an α-amino group but not having an ε-amino group and the other peptide fragments (B) having an α-amino group and an ε-amino group, selectively modifying the α-amino groups to obtain a C-terminal peptide fragment modified (A′) and the other peptide fragments modified (B′); and separating the C-terminal peptide fragment modified (A′) from the modified cleavage product by allowing a carrier to hold the other peptide fragments modified (B′) via the ε-amino group. A method for determining the amino acid sequence of a C-terminal peptide of a protein, comprising the steps of: selectively recovering a C-terminal peptide of a protein by the above method; and determining the amino acid sequence by subjecting a recovered C-terminal peptide fragment to mass spectrometry measurement.

摘要翻译： 本发明提供了特异性回收C末端肽片段的方法，以及通过使用质谱仪容易地测定难以用常规方法测定的C末端肽片段的序列的方法 ，特别是能够重新测序C-末端肽片段的方法。 一种选择性回收蛋白质C末端肽的方法，包括以下步骤：在含有具有α-氨基但不具有ε-氨基的C-末端肽片段（A）的蛋白质的切割产物中 和具有α-氨基和ε-氨基的其它肽片段（B），选择性地修饰α-氨基以获得修饰的C末端肽片段（A'）和修饰的其它肽片段（B' ）; 以及通过允许载体通过ε-氨基保持修饰的其它肽片段（B'），从修饰的切割产物中分离修饰的C末端肽片段（A'）。 一种确定蛋白质C-末端肽的氨基酸序列的方法，包括以下步骤：通过上述方法选择性地回收蛋白质的C-末端肽; 并通过对回收的C-末端肽片段进行质谱测量来测定氨基酸序列。