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公开(公告)号：US09518081B2
公开(公告)日：2016-12-13
申请号：US13390252
申请日：2010-08-12
CPC分类号： C12N15/1037 , C07K1/00 , C07K1/042 , C07K1/113 , C07K2318/00
摘要： The invention relates to a method for altering the conformational diversity of a first repertoire of polypeptide ligands, comprising a plurality of polypeptides comprising at least two reactive groups separated by a loop sequence covalently linked to a molecular scaffold which forms covalent bonds with said reactive groups, to produce a second repertoire of polypeptide ligands, comprising assembling said second repertoire from the polypeptides and structural scaffold of said first repertoire, incorporating one of the following alterations: (a) altering at least one reactive group; or (b) altering the nature of the molecular scaffold; or (c) altering the bond between at least one reactive group and the molecular scaffold; or (d) any combination of (a), (b) or (c).
摘要翻译： 本发明涉及一种用于改变多肽配体的第一组分的构象多样性的方法，其包含多个多肽，所述多肽包含至少两个反应性基团，所述反应基团通过共价连接到与所述反应性基团形成共价键的分子支架的环序列分开， 以产生多肽配体的第二组成成分，其包括从所述第一组合物的多肽和结构性支架组装所述第二组分，并结合以下改变之一：（a）改变至少一个反应性基团; 或（b）改变分子支架的性质; 或（c）改变至少一个反应性基团和分子支架之间的键; 或（d）（a），（b）或（c）的任何组合。
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公开(公告)号：US20120172235A1
公开(公告)日：2012-07-05
申请号：US13390252
申请日：2010-08-12
CPC分类号： C12N15/1037 , C07K1/00 , C07K1/042 , C07K1/113 , C07K2318/00
摘要： The invention relates to a method for altering the conformational diversity of a first repertoire of polypeptide ligands, comprising a plurality of polypeptides comprising at least two reactive groups separated by a loop sequence covalently linked to a molecular scaffold which forms covalent bonds with said reactive groups, to produce a second repertoire of polypeptide ligands, comprising assembling said second repertoire from the polypeptides and structural scaffold of said first repertoire, incorporating one of the following alterations: (a) altering at least one reactive group; or (b) altering the nature of the molecular scaffold; or (c) altering the bond between at least one reactive group and the molecular scaffold; or (d) any combination of (a), (b) or (c).
摘要翻译： 本发明涉及一种用于改变多肽配体的第一组分的构象多样性的方法，其包含多个多肽，所述多肽包含至少两个反应性基团，所述反应基团通过共价连接到与所述反应性基团形成共价键的分子支架的环序列分开， 以产生多肽配体的第二组成成分，其包括从所述第一组合物的多肽和结构性支架组装所述第二组分，并结合以下改变之一：（a）改变至少一个反应性基团; 或（b）改变分子支架的性质; 或（c）改变至少一个反应性基团和分子支架之间的键; 或（d）（a），（b）或（c）的任何组合。
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公开(公告)号：US08685890B2
公开(公告)日：2014-04-01
申请号：US13147289
申请日：2010-02-04
申请人： Gregory Paul Winter , Christian Heinis , Elise Bernard , David Loakes , John Tite , Marina Vaysburd , Daniel Paul Teufel , Lutz Riechmann
发明人： Gregory Paul Winter , Christian Heinis , Elise Bernard , David Loakes , John Tite , Marina Vaysburd , Daniel Paul Teufel , Lutz Riechmann
IPC分类号： C40B30/04
CPC分类号： C12N15/1037 , A61K47/55 , A61K47/64 , A61K47/643 , C07K14/001 , C12N15/1044 , C12N15/1058 , C40B40/08 , C40B40/10 , C40B50/06 , G01N33/531
摘要： The invention relates to a method for providing a multispecific peptide ligand comprising a polypeptide covalently linked to a molecular scaffold at three or more amino acid residues and capable of binding to two or more separate targets, comprising the steps of: (a) providing a first repertoire of polypeptides, each polypeptide comprising two or more reactive groups capable of covalent linkage to a molecular scaffold, and at least one loop which comprises a sequence of two or more amino acids subtended between two of said reactive groups; (b) providing a second repertoire of polypeptides as described in (a); (c) joining at least one loop of one or more members of the first repertoire to at least one loop of one or more members of the second repertoire to form at least one polypeptide comprising two loops, and (d) conjugating the composite polypeptide(s) to a molecular scaffold at at least three amino acid positions.
摘要翻译： 本发明涉及一种提供多特异性肽配体的方法，所述多特异性肽配体包含在三个或更多个氨基酸残基处共价连接到分子支架并能够结合两个或更多个单独靶标的多肽，包括以下步骤：（a）提供第一 多肽的所有组成部分，每个多肽包含两个或更多个能够与分子支架共价连接的反应性基团，以及至少一个环，其包含两个或更多个氨基酸的序列，对应于两个所述反应性基团; （b）提供如（a）所述的第二组多肽; （c）将第一组合物中的一个或多个成员的至少一个环连接到第二组合物的一个或多个成员的至少一个环，以形成至少一个包含两个环的多肽，和（d）将复合多肽（ s）至少在三个氨基酸位置分子支架。
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公开(公告)号：US20120101256A1
公开(公告)日：2012-04-26
申请号：US13147289
申请日：2010-02-04
申请人： Gregory Paul Winter , Christian Heinis , Elise Bernard , David Loakes , John Tite , Marina Vaysburd , Daniel Paul Teufel , Lutz Riechmann
发明人： Gregory Paul Winter , Christian Heinis , Elise Bernard , David Loakes , John Tite , Marina Vaysburd , Daniel Paul Teufel , Lutz Riechmann
CPC分类号： C12N15/1037 , A61K47/55 , A61K47/64 , A61K47/643 , C07K14/001 , C12N15/1044 , C12N15/1058 , C40B40/08 , C40B40/10 , C40B50/06 , G01N33/531
摘要： The invention relates to a method for providing a multispecific peptide ligand comprising a polypeptide covalently linked to a molecular scaffold at three or more amino acid residues and capable of binding to two or more separate targets, comprising the steps of: (a) providing a first repertoire of polypeptides, each polypeptide comprising two or more reactive groups capable of covalent linkage to a molecular scaffold, and at least one loop which comprises a sequence of two or more amino acids subtended between two of said reactive groups; (b) providing a second repertoire of polypeptides as described in (a); (c) joining at least one loop of one or more members of the first repertoire to at least one loop of one or more members of the second repertoire to form at least one polypeptide comprising two loops, and (d) conjugating the composite polypeptide(s) to a molecular scaffold at at least three amino acid positions.
摘要翻译： 本发明涉及一种提供多特异性肽配体的方法，所述多特异性肽配体包含在三个或更多个氨基酸残基处共价连接到分子支架并能够结合两个或更多个单独靶标的多肽，包括以下步骤：（a）提供第一 多肽的所有组成部分，每个多肽包含两个或更多个能够与分子支架共价连接的反应性基团，以及至少一个环，其包含两个或更多个氨基酸的序列，对应于两个所述反应性基团; （b）提供如（a）所述的第二组多肽; （c）将第一组合物中的一个或多个成员的至少一个环连接到第二组合物的一个或多个成员的至少一个环，以形成至少一个包含两个环的多肽，和（d）将复合多肽（ s）至少在三个氨基酸位置分子支架。
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公开(公告)号：US08778844B2
公开(公告)日：2014-07-15
申请号：US13147257
申请日：2010-02-04
IPC分类号： C40B30/04
CPC分类号： C12N15/1037 , A61K47/55 , A61K47/64 , A61K47/643 , C07K14/001 , C12N15/1044 , C12N15/1058 , C40B40/08 , C40B40/10 , C40B50/06 , G01N33/531
摘要： The invention relates to a method for modifying one or more peptide ligands, comprising polypeptides covalently linked to a molecular scaffold at two or more amino acid residues, comprising the steps of providing one or more peptide ligands, wherein the polypeptide comprises two or more reactive groups which form a covalent linkage to the molecular scaffold, and at least one loop which comprises a sequence of two or more amino acids subtended between two of said reactive groups; exposing the peptide ligands to one or more proteases; and sorting the ligands according to the extent of proteolytic cleavage.
摘要翻译： 本发明涉及一种修饰一种或多种肽配体的方法，其包括在两个或更多个氨基酸残基处与分子支架共价连接的多肽，包括提供一个或多个肽配体的步骤，其中所述多肽包含两个或多个反应性基团 其形成与分子支架的共价键，以及至少一个环，其包含在两个所述反应性基团之间对位的两个或更多个氨基酸的序列; 将肽配体暴露于一种或多种蛋白酶; 并根据蛋白水解切割的程度对配体进行分选。
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公开(公告)号：US20120142541A1
公开(公告)日：2012-06-07
申请号：US13147257
申请日：2010-02-04
CPC分类号： C12N15/1037 , A61K47/55 , A61K47/64 , A61K47/643 , C07K14/001 , C12N15/1044 , C12N15/1058 , C40B40/08 , C40B40/10 , C40B50/06 , G01N33/531
摘要： The invention relates to a method for modifying one or more peptide ligands, comprising polypeptides covalently linked to a molecular scaffold at two or more amino acid residues, comprising the steps of providing one or more peptide ligands, wherein the polypeptide comprises two or more reactive groups which form a covalent linkage to the molecular scaffold, and at least one loop which comprises a sequence of two or more amino acids subtended between two of said reactive groups; exposing the peptide ligands to one or more proteases; and sorting the ligands according to the extent of proteolytic cleavage.
摘要翻译： 本发明涉及一种修饰一种或多种肽配体的方法，其包括在两个或更多个氨基酸残基处与分子支架共价连接的多肽，包括提供一个或多个肽配体的步骤，其中所述多肽包含两个或多个反应性基团 其形成与分子支架的共价键，以及至少一个环，其包含在两个所述反应性基团之间对位的两个或更多个氨基酸的序列; 将肽配体暴露于一种或多种蛋白酶; 并根据蛋白水解切割的程度对配体进行分选。
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公开(公告)号：US20120101253A1
公开(公告)日：2012-04-26
申请号：US13147308
申请日：2010-02-04
IPC分类号： C07K7/64
CPC分类号： C12N15/1037 , A61K47/55 , A61K47/64 , A61K47/643 , C07K14/001 , C12N15/1044 , C12N15/1058 , C40B40/08 , C40B40/10 , C40B50/06 , G01N33/531
摘要： The invention relates to a peptide ligand comprising a polypeptide linked to a molecular scaffold at n attachment points, wherein said polypeptide is cyclised and forms n separate loops subtended between said n attachment points on the molecular scaffold, wherein n is greater than or equal to 2.
摘要翻译： 本发明涉及一种肽配体，其包含在n个连接点处与分子支架连接的多肽，其中所述多肽被环化并形成对分子支架上的所述n个附着点之间的n个分开的环，其中n大于或等于2 。
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公开(公告)号：US20070105117A1
公开(公告)日：2007-05-10
申请号：US10574227
申请日：2004-10-01
申请人： Christian Heinis
发明人： Christian Heinis
CPC分类号： C12N15/1075
摘要： The invention relates to a method for the production and allocation of nucleic acids and polypeptides coded thereby. Said method can be used for evolutionary selection of polypeptides in vitro. The inventive method makes it possible to allocate nucleic acids to the polypeptides coded thereby and to select and isolate nucleic acids coding for polypeptides with selected properties. The invention is directed to the use of (cytosin-5)-methyl transferases and to the use of fusion polypeptides or nucleic acid fusion polypeptide complexes covalently bonded thereto according to the inventive method.
摘要翻译： 本发明涉及一种生产和分配由其编码的核酸和多肽的方法。 所述方法可用于体外进化多肽的选择。 本发明的方法使得可以将核酸分配给由其编码的多肽，并选择和分离编码具有选定性质的多肽的核酸。 本发明涉及根据本发明方法使用（细胞因子-5） - 甲基转移酶和共价共价键合的融合多肽或核酸融合多肽复合物。
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公开(公告)号：US20110165593A1
公开(公告)日：2011-07-07
申请号：US13006087
申请日：2011-01-13
申请人： Jan Barnikow , Christopher Chidley , Thomas Gronemeyer , Christian Heinis , Hughes Jaccard , Kai Johnsson , Alexandre Juillerat , Antje Keppler
发明人： Jan Barnikow , Christopher Chidley , Thomas Gronemeyer , Christian Heinis , Hughes Jaccard , Kai Johnsson , Alexandre Juillerat , Antje Keppler
IPC分类号： G01N33/573 , C12N9/10
CPC分类号： C12N9/10 , C07K2319/00 , C12N9/1007
摘要： The invention relates to AGT mutants showing, when compared to the wild type human AGT, two or more advantageous properties selected from (a) reduced DNA interaction; (b) localisation of the expressed protein in eukaryotic cells that is no longer restricted to the nucleus; (c) improved expression yield as soluble protein and improved stability in various hosts; (d) improved stability under oxidising conditions; (e) improved stability within cells after reaction with a substrate; (f) improved stability outside cells before and after reaction with a substrate; (g) improved in vitro solubility; (h) improved reactivity against O6-alkylguanine substrates; (i) reduced reactivity against DNA-based substrates; and (j) reduced reactivity against N9-substituted O6-alkylguanine substrates. Such AGT mutants with the mentioned improved properties are mutants wherein between 1 and 25 amino acids of the wild type human AGT are substituted by other amino acids, and optionally 1 to 5 amino acids out of the continuous chain at one, two or three positions are deleted or added and/or 1 to 4 amino acids at the N-terminus or 1 to 40 amino acids at the C-terminus are deleted. The invention further relates to a method for detecting and/or manipulating a protein of interest wherein the protein of interest is incorporated into a fusion protein with the AGT mutants of the invention. Another object of the invention are AGT fusion proteins comprising such AGT mutants and the protein of interest.
摘要翻译： 本发明涉及AGT突变体，当与野生型人AGT相比时，显示出选自（a）减少的DNA相互作用的两种或更多种有利性质; （b）表达的蛋白质在不再限于细胞核的真核细胞中的定位; （c）提高作为可溶性蛋白质的表达产率和改善各种宿主的稳定性; （d）改善氧化条件下的稳定性; （e）在与底物反应后改善细胞内的稳定性; （f）在与底物反应之前和之后提高细胞外的稳定性; （g）提高体外溶解度; （h）改善对O6-烷基鸟嘌呤底物的反应性; （i）降低对DNA基底层的反应性; 和（j）对N9取代的O6-烷基鸟嘌呤底物的反应性降低。 具有上述改进性质的这种AGT突变体是突变体，其中野生型人AGT的1至25个氨基酸被其它氨基酸取代，并且在一个，两个或三个位置上连续链中任选地1至5个氨基酸是 在N-末端缺失或添加和/或1至4个氨基酸或C末端的1至40个氨基酸被缺失。 本发明还涉及用于检测和/或操纵感兴趣的蛋白质的方法，其中将目的蛋白质与本发明的AGT突变体掺入融合蛋白中。 本发明的另一个目的是包含这种AGT突变体和目的蛋白质的AGT融合蛋白。
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公开(公告)号：US07527954B2
公开(公告)日：2009-05-05
申请号：US10574227
申请日：2004-10-01
CPC分类号： C12N15/1075
摘要： The invention relates to a method for the production and allocation of nucleic acids and polypeptides coded thereby. The method can be used for evolutionary selection of polypeptides in vitro. The inventive method makes it possible to allocate nucleic acids to the polypeptides coded thereby and to select and isolate nucleic acids coding for polypeptides with selected properties. The invention is directed to the use of (cytosin-5)-methyl transferases and to the use of fusion polypeptides or nucleic acid fusion polypeptide complexes covalently bonded thereto according to the inventive method.
摘要翻译： 本发明涉及一种生产和分配由其编码的核酸和多肽的方法。 该方法可用于体外进化多肽的选择。 本发明的方法使得可以将核酸分配给由其编码的多肽，并选择和分离编码具有选定性质的多肽的核酸。 本发明涉及根据本发明方法使用（细胞因子-5） - 甲基转移酶和共价共价键合的融合多肽或核酸融合多肽复合物。
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