Site-specific labelling of proteins using acridone and quinacridone lifetime dyes
    3.
    发明申请
    Site-specific labelling of proteins using acridone and quinacridone lifetime dyes 审中-公开
    使用吖啶酮和喹吖啶酮终身染料的蛋白质的位点特异性标记

    公开(公告)号:US20060051811A1

    公开(公告)日:2006-03-09

    申请号:US10522665

    申请日:2003-07-28

    申请人: Graham Cotton

    发明人: Graham Cotton

    摘要: Disclosed are compounds of formula: in which D is a fluorescent dye selected from an acridone and a quinacridone dye; F comprises a target bonding group selected from a carboxylic acid thioester group and a 1,2-aminothiol group; M is a group adapted for attaching to F; X is selected from hydrogen or the group: -L2-B wherein B is an affinity tag; and L1 and L2 each independently comprise a group containing from 1 to 40 linked atoms selected from carbon atoms which may optionally include one or more groups selected from —NR′—, —O—, —CH═CH—, —CO—NH— and phenylenyl groups, where R′ is selected from hydrogen and C1-C4 alkyl. The invention also relates to methods that afford direct attachment of the acridone or quinacridone dye to either the N-terminus or C-terminus of a synthetic or recombinant peptide or protein, and their derivatives, in a site-specific manner, coupled with purification of the resultant labelled molecule.

    摘要翻译: 公开了下式的化合物:

    METHOD OF LABELLING INTERFERONS WITH PEG
    4.
    发明申请
    METHOD OF LABELLING INTERFERONS WITH PEG 审中-公开
    用PEG标记干扰素的方法

    公开(公告)号:US20120128629A1

    公开(公告)日:2012-05-24

    申请号:US13265032

    申请日:2010-05-17

    CPC分类号: C07K14/56 A61K38/00 A61K47/60

    摘要: A method of site specific labelling of an interferon molecule is provided. The method comprises the steps: a) providing a label molecule comprising a PEG moiety having an aldehyde or ketone moiety; b) providing an interferon molecule having a C terminal hydrazide moiety; and c) allowing the aldehyde or ketone moiety of the PEG moiety to react with the C terminal hydrazide of the interferon molecule to form a labelled interferon molecule, which comprises a PEG moiety attached to the C terminus of the interferon molecule via a hydrazone bond. Interferon molecules labelled using such a method are also described.

    摘要翻译: 提供了干扰素分子的位点特异性标记方法。 该方法包括以下步骤:a)提供包含具有醛或酮部分的PEG部分的标记分子; b)提供具有C末端酰肼部分的干扰素分子; 和c)允许PEG部分的醛或酮部分与干扰素分子的C末端酰肼反应以形成标记的干扰素分子,其包含通过腙键连接至干扰素分子的C末端的PEG部分。 还描述了使用这种方法标记的干扰素分子。

    Ligation method
    5.
    发明授权
    Ligation method 有权
    连接法

    公开(公告)号:US07622552B2

    公开(公告)日:2009-11-24

    申请号:US10567403

    申请日:2004-08-05

    申请人: Graham Cotton

    发明人: Graham Cotton

    IPC分类号: C07K1/08 C07K1/107 C07K2/00

    CPC分类号: C07K1/1077

    摘要: A method of ligating two or more molecules, for example, peptides to peptides or peptides to labels is provided. The method may comprise the steps: a) providing a first oligopeptide having a reactive moiety, which is a hydrazine moiety, a hydrazide moiety or an amino-óxy moiety b) providing a second oligopeptide having an activated ester moiety, c)allowing the reactive moiety of the first oligopeptide to react with the activated ester moiety of the second oligopeptide to form an oligopeptide product, in which the first and second oligopeptides are linked via a linking moiety having Formula I, II or III. The second oligopeptide may preferably be generated by thiol reagent induced cleavage of an intein fusion protein. The invention further provides labelling and ligation methods in which protein hydrazides are ligated by reaction of the hydrazide moiety with an aldehyde functionality or a ketone functionality.

    摘要翻译: 提供了将两个或多个分子例如肽与肽或肽连接至标记的方法。 该方法可以包括以下步骤:a)提供具有反应性部分的第一寡肽,其为肼部分,酰肼部分或氨基-o-部分b)提供具有活化酯部分的第二寡肽,c)允许反应性 第一寡肽的部分与第二寡肽的活化酯部分反应以形成寡肽产物,其中第一和第二寡肽通过具有式I,II或III的连接部分连接。 第二寡肽可以优选地由硫醇试剂诱导的内蛋白融合蛋白的切割产生。 本发明进一步提供了标记和连接方法,其中蛋白质酰肼通过酰肼部分与醛官能团或酮官能团的反应连接。

    Purification means
    7.
    发明申请
    Purification means 审中-公开
    净化手段

    公开(公告)号:US20070010673A1

    公开(公告)日:2007-01-11

    申请号:US10558870

    申请日:2004-06-01

    IPC分类号: C07D401/00

    CPC分类号: C07K1/13 C07D213/38

    摘要: Disclosed is a tag for purification of peptides. The tag structure facilitates the easy cleavage of a bond formed between the tag molecule and a peptide to which it is bound under conditions which minimize, indeed preferably prevent, damage to the peptide. The tag molecules of the invention may be used to separate and/or purify a peptide from a mixture of peptides and/or other components.

    摘要翻译: 公开了用于纯化肽的标签。 在最小化,实际上优选地防止对肽损伤的条件下,标签结构有助于容易地切割标签分子与其所结合的肽之间形成的键。 本发明的标签分子可用于从肽和/或其它组分的混合物分离和/或纯化肽。

    Ligation method
    8.
    发明申请

    公开(公告)号:US20060247417A1

    公开(公告)日:2006-11-02

    申请号:US10567403

    申请日:2004-08-05

    申请人: Graham Cotton

    发明人: Graham Cotton

    IPC分类号: A61K38/16 C07K7/08 C07K1/02

    CPC分类号: C07K1/1077

    摘要: A method of ligating two or more molecules, for example, peptides to peptides or peptides to labels is provided. The method may comprise the steps: a) providing a first oligopeptide having a reactive moiety, which is a hydrazine moiety, a hydrazide moiety or an amino-óxy moiety b) providing a second oligopeptide having an activated ester moiety, c)allowing the reactive moiety of the first oligopeptide to react with the activated ester moiety of the second oligopeptide to form an oligopeptide product, in which the first and second oligopeptides are linked via a linking moiety having Formula I, II or III. The second oligopeptide may preferably be generated by thiol reagent induced cleavage of an intein fusion protein. The invention further provides labelling and ligation methods in which protein hydrazides are ligated by reaction of the hydrazide moiety with an aldehyde functionality or a ketone functionality.

    Multiple sensor-containing active modified polypeptides, preparation and uses thereof
    9.
    发明申请
    Multiple sensor-containing active modified polypeptides, preparation and uses thereof 审中-公开
    含多个含传感器的活性修饰的多肽,其制备和用途

    公开(公告)号:US20060040319A1

    公开(公告)日:2006-02-23

    申请号:US10784721

    申请日:2004-02-23

    IPC分类号: G01N33/53

    摘要: Compositions and methods are provided for identifying conformational changes in polypeptides related to the activity or biological state of the polypeptide. Semisynthetic polypeptides are prepared comprising at least two proximity-sensor peptides, the resultant composition capable of detectably indicating the activity of biological state of the polypeptide. Such compositions may be used to identify modulators of the polypeptides as well as modulators of molecules which interact with the polypeptide, such as protein kinases which act on protein kinase targets.

    摘要翻译: 提供组合物和方法用于鉴定与多肽的活性或生物学状态相关的多肽中的构象变化。 制备包含至少两个接近传感器肽的半合成多肽,所得组合物能够可检测地指示多肽的生物学状态的活性。 这样的组合物可用于鉴定多肽的调节剂以及与多肽相互作用的分子调节剂,例如作用于蛋白激酶靶标的蛋白激酶。

    Site-specific labelling of proteins using cyanine dye reporters
    10.
    发明申请
    Site-specific labelling of proteins using cyanine dye reporters 审中-公开
    使用花青染料记者对蛋白质的位点特异性标记

    公开(公告)号:US20050239144A1

    公开(公告)日:2005-10-27

    申请号:US10522675

    申请日:2003-07-28

    申请人: Graham Cotton

    发明人: Graham Cotton

    摘要: Disclosed are compounds of formula: in which D is a dye selected from a cyanine dye or a derivative thereof; B is an affinity tag; F comprises a target bonding group selected from a carboxylic acid thioester group and a 1,2-aminothiol group; M is a group adapted for attaching to F; and L1 and L2 each independently comprise a group containing from 1 to 40 linked atoms selected from carbon atoms which may optionally include one or more groups selected from —NR′—, —O—, —CH═CH—, —CO—NH— and phenylenyl groups, where R′ is selected from hydrogen and C1-C4 alkyl. The invention also relates to methods that afford direct attachment of the cyanine dye reporter group to either the N-terminus or C-terminus of a synthetic or recombinant peptide or protein, and their derivatives, in a site-specific manner, coupled with purification of the resultant labelled molecule.

    摘要翻译: 公开了下式的化合物:其中D是选自花青染料或其衍生物的染料; B是亲和标签; F包含选自羧酸硫酯基和1,2-氨基硫醇基的目标键合基团; M是适合附着于F的组; 和L 1和L 2各自独立地包含含有1至40个选自碳原子的连接原子的基团,其可任选地包括一个或多个选自-NR'- ,-O - , - CH-CH - , - CO-NH-和苯基,其中R'选自氢和C 1 -C 4烷基。 本发明还涉及将花青染料报告基团直接连接到合成或重组肽或蛋白质的N末端或C末端的方法及其衍生物,以位点特异性方式与纯化 得到的标记分子。