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1. 发明申请

US20050208577A1 In vivo library-versus-library selection of optimized protein-protein interactions 失效
标题翻译：体内文库对文库选择优化的蛋白质 - 蛋白质相互作用
公开(公告)号：US20050208577A1
公开(公告)日：2005-09-22
申请号：US11134253
申请日：2005-05-23
申请人： Stephen Michnick , Joelle Pelletier , Katja Arndt , Andreas Pluckthun
发明人： Stephen Michnick , Joelle Pelletier , Katja Arndt , Andreas Pluckthun
IPC分类号： C07K14/435 , C12N15/10 , C12Q1/32 , C12Q1/68 , C40B30/04 , G01N33/53 , G01N33/542 , G01N33/573 , G01N33/58 , G01N33/68
CPC分类号： C40B30/04 , C07K14/43595 , C07K2319/00 , C12N15/1055 , C12Q1/32 , G01N33/542 , G01N33/6845 , Y02A90/26
摘要： The present invention describes a rapid and efficient in vivo library-versus-library screening strategy for identifying optimally interacting pairs of heterodimerizing polypeptides. It allows for the screening of a protein library against a second protein library, rather than against a single bait protein, and thus has numerous applications in the study of protein-protein interactions. Additionally, it allows for the application of different selection stringencies. Two leucine zipper libraries, semi-randomized at the positions adjacent to the hydrophobic core, were genetically fused to either one of two designed fragments of the enzyme murine dihydrofolate reductase (mDHFR), and cotransformed into E. coli. Interaction between the library polypeptides was required for reconstitution of the enzymatic activity of mDHFR, allowing bacterial growth. Analysis of the resulting colonies revealed important biases in the zipper sequences relative to the original libraries, which are consistent with selection for stable, heterodimerizing pairs. Using more weakly associating mDHFR fragments, we increased the stringency of selection. We enriched the best performing leucine zipper pairs by multiple passaging of the pooled, selected colonies in liquid culture, as the best pairs allowed for better bacterial propagation. This competitive growth allowed small differences among the pairs to be amplified, and different sequence positions were enriched at different rates. We applied these selection processes to a library-versus-library sample of 2.0×106 combinations, and selected a novel leucine zipper pair which may be appropriate for use in further in vivo heterodimerization strategies.
摘要翻译：本发明描述了快速和有效的体内文库对文库筛选策略，用于鉴定异构二聚化多肽的最佳相互作用对。它允许针对第二蛋白质文库筛选蛋白质文库，而不是针对单个诱饵蛋白质，因此在蛋白质 - 蛋白质相互作用的研究中具有许多应用。此外，它允许应用不同的选择严格性。两个亮氨酸拉链文库在与疏水核心相邻的位置半随机化，与酶二氢叶酸还原酶（mDHFR）的两个设计的片段中的任一个遗传融合，并共转化到大肠杆菌中。需要文库多肽之间的相互作用来重构mDHFR的酶活性，从而允许细菌生长。所得菌落的分析揭示了相对于原始文库的拉链序列的重要偏倚，这与选择稳定的异源二聚体对一致。使用更弱的关联mDHFR片段，我们增加了选择的严格性。我们通过在液体培养中多次传代合并的选定菌落，丰富了表现最好的亮氨酸拉链对，因为最佳对允许更好的细菌繁殖。这种竞争性增长允许放大对中的小差异，并且以不同的速率富集不同的序列位置。我们将这些选择过程应用于2.0×10 6组合的文库对文库样品，并选择了可能适用于进一步体内异二聚化策略的新型亮氨酸拉链对。

2. 发明申请

US20120142611A1 REPEAT PROTEIN FROM COLLECTION OF REPEAT PROTEINS COMPRISING REPEAT MODULES 有权
标题翻译：收集包含重复模块的重复蛋白质的蛋白质
公开(公告)号：US20120142611A1
公开(公告)日：2012-06-07
申请号：US13330336
申请日：2011-12-19
申请人： Michael Tobias STUMPP , Patrik Forrer , Hans Kaspar Binz , Andreas Pluckthun
发明人： Michael Tobias STUMPP , Patrik Forrer , Hans Kaspar Binz , Andreas Pluckthun
IPC分类号： A61K38/16 , C07K14/00 , C07H21/00 , C12N15/63
CPC分类号： C12N15/1055 , A61K38/00 , A61K38/16 , C07K14/00 , C07K14/47 , C07K14/4703 , C12N15/10 , C12N15/1037 , C12N15/1044 , C12N15/1093
摘要： A repeat protein from a collection of repeat proteins, wherein each repeat protein of said collection comprises a repeat domain, which comprises a set of consecutive repeat modules, wherein the repeat modules have the same fold and stack tightly to create a superhelical structure having a joint hydrophobic core, wherein each of the repeat modules is derived from one or more repeat units and wherein the repeat units comprise framework residues, which contribute to the folding topology of the repeat unit or contribute to an interaction with a neighboring repeat unit, and target interaction residues, which contribute to an interaction with a target substance, wherein the repeat proteins of the collection differ from other repeat proteins in the collection in at least one amino acid position of the repeat modules is described as are related pharmaceuticals and nucleic acid molecules.
摘要翻译：来自重复蛋白质的重复蛋白质，其中所述收集物的每个重复蛋白质包含重复结构域，其包含一组连续重复模块，其中所述重复模块具有相同的折叠并且紧密堆叠以产生具有关节的超螺旋结构疏水核心，其中每个重复模块衍生自一个或多个重复单元，并且其中重复单元包括框架残基，其有助于重复单元的折叠拓扑或有助于与相邻重复单元的相互作用，以及靶相互作用有助于与靶物质相互作用的残基，其中所述集合的重复蛋白质与所述重复模块的至少一个氨基酸位置中的所述集合中的其它重复蛋白质不同描述为相关药物和核酸分子。

3. 发明授权

US06897017B1 Vivo library-versus-library selection of optimized protein-protein interactions 失效
标题翻译：体内文库与图书馆选择优化的蛋白质 - 蛋白质相互作用
公开(公告)号：US06897017B1
公开(公告)日：2005-05-24
申请号：US09603885
申请日：2000-06-26
申请人： Stephen William Watson Michnick , Joelle N. Pelletier , Katja M. Arndt , Andreas Pluckthun
发明人： Stephen William Watson Michnick , Joelle N. Pelletier , Katja M. Arndt , Andreas Pluckthun
IPC分类号： C07K14/435 , C12N15/10 , C12Q1/32 , C12Q1/68 , C40B30/04 , G01N33/53 , G01N33/542 , G01N33/573 , G01N33/58 , G01N33/68 , C12N15/52 , C12P21/02
CPC分类号： C40B30/04 , C07K14/43595 , C07K2319/00 , C12N15/1055 , C12Q1/32 , G01N33/542 , G01N33/6845 , Y02A90/26
摘要： The present invention describes a rapid and efficient in vivo library-versus-library screening strategy for identifying optimally interacting pairs of heterodimerizing polypeptides. It allows for the screening of a protein library against a second protein library, rather than against a single bait protein, and thus has numerous applications in the study of protein-protein interactions. Additionally, it allows for the application of different selection stringencies. Two leucine zipper libraries, semi-randomized at the positions adjacent to the hydrophobic core, were genetically fused to either one of two designed fragments of the enzyme murine dihydrofolate reductase (mDHFR), and cotransformed into E. coli. Interaction between the library polypeptides was required for reconstitution of the enzymatic activity of mDHFR, allowing bacterial growth. Analysis of the resulting colonies revealed important biases in the zipper sequences relative to the original libraries, which are consistent with selection for stable, heterodimerizing pairs. Using more weakly associating mDHFR fragments, we increased the stringency of selection. We enriched the best performing leucine zipper pairs by multiple passaging of the pooled, selected colonies in liquid culture, as the best pairs allowed for better bacterial propagation. This competitive growth allowed small differences among the pairs to be amplified, and different sequence positions were enriched at different rates. We applied these selection processes to a library-versus-library sample of 2.0×106 combinations, and selected a novel leucine zipper pair which may be appropriate for use in further in vivo heterodimerization strategies.
摘要翻译：本发明描述了快速和有效的体内文库对文库筛选策略，用于鉴定异构二聚化多肽的最佳相互作用对。它允许针对第二蛋白质文库筛选蛋白质文库，而不是针对单个诱饵蛋白质，因此在蛋白质 - 蛋白质相互作用的研究中具有许多应用。此外，它允许应用不同的选择严格性。两个亮氨酸拉链文库在与疏水核心相邻的位置半随机化，与酶二氢叶酸还原酶（mDHFR）的两个设计的片段中的任一个遗传融合，并共转化到大肠杆菌中。需要文库多肽之间的相互作用来重构mDHFR的酶活性，从而允许细菌生长。所得菌落的分析揭示了相对于原始文库的拉链序列的重要偏倚，这与选择稳定的异源二聚体对一致。使用更弱的关联mDHFR片段，我们增加了选择的严格性。我们通过在液体培养中多次传代合并的选定菌落，丰富了表现最好的亮氨酸拉链对，因为最佳对允许更好的细菌繁殖。这种竞争性增长允许放大对中的小差异，并且以不同的速率富集不同的序列位置。我们将这些选择过程应用于2.0×10 6组合的文库对文库样品，并选择了可能适用于进一步体内异二聚化策略的新型亮氨酸拉链对。

4. 发明授权

US5514548A Method for in vivo selection of ligand-binding proteins 失效
标题翻译：体内选择配体结合蛋白的方法
公开(公告)号：US5514548A
公开(公告)日：1996-05-07
申请号：US197770
申请日：1994-02-17
申请人： Klaus Krebber , Simon Moroney , Andreas Pluckthun , Christian Schneider
发明人： Klaus Krebber , Simon Moroney , Andreas Pluckthun , Christian Schneider
IPC分类号： C12N1/21 , C12N7/00 , C12N15/09 , C12N15/10 , C12Q1/68 , C12Q1/70 , C40B40/02 , G01N33/53 , G01N33/566 , C12N15/64 , C12N15/70
CPC分类号： C40B40/02 , C12N15/1037 , C12Q1/686 , C12Q1/70
摘要： The application discloses a method, and kit, which enables the selection of proteins or other oligo- or polypeptides (collectively, ligand binding proteins, LBPs) which bind with high affinity to a target ligand or receptor. The method relies on the display of the LBP of interest on the surface of replicable genetic packages (RGPs) which are modified so as to be non-infective. Infectivity is restored to those RGPs displaying LBPs with high affinity for a target ligand or receptor by an infectivity mediating complex. The infectivity mediating complex comprises the ligand or receptor covalently linked to a polypeptide which brings about interaction between the RGP and a host cell. It is envisaged that the LBP of interest will belong to a genetically diverse collection of similar substances. The method therefore allows selection of substances with high affinity for target, from within a large collection of variants.
摘要翻译：该申请公开了一种方法和试剂盒，其能够选择以高亲和力结合至靶配体或受体的蛋白质或其它寡聚物或多肽（统称为配体结合蛋白，LBP）。该方法依赖于在可复制遗传包（RGP）的表面上显示感兴趣的LBP，其被修饰为非感染性。通过传染性介导复合物，对目标配体或受体具有高亲和力的显示LBP的那些RGP恢复感染性。感染性介导复合物包含与导致RGP和宿主细胞之间的相互作用的多肽共价连接的配体或受体。预期感兴趣的LBP将属于类似物质的遗传多样性收集。因此，该方法允许从大量变体中选择对目标具有高亲和力的物质。

5. 发明授权

US08110653B2 Collections of repeat proteins comprising repeat modules 有权
标题翻译：包含重复模块的重复蛋白质的集合
公开(公告)号：US08110653B2
公开(公告)日：2012-02-07
申请号：US12222525
申请日：2008-08-11
申请人： Michael Tobias Stumpp , Patrik Forrer , Hans Kasper Binz , Andreas Pluckthun
发明人： Michael Tobias Stumpp , Patrik Forrer , Hans Kasper Binz , Andreas Pluckthun
IPC分类号： A61K38/00 , C07K14/00
CPC分类号： C12N15/1055 , A61K38/00 , A61K38/16 , C07K14/00 , C07K14/47 , C07K14/4703 , C12N15/10 , C12N15/1037 , C12N15/1044 , C12N15/1093
摘要： A collection of repeat proteins, each repeat protein comprising a repeat domain, which comprises a set of consecutive repeat modules, wherein each of the repeat modules is derived from one or more repeat units and wherein the repeat units comprise framework residues, which contribute to the folding topology of the repeat unit or contribute to an interaction with a neighboring repeat unit, and target interaction residues, which contribute to an interaction with a target substance, wherein the repeat proteins differ from other repeat proteins in the collection in at least one amino acid position of the repeat modules is described.
摘要翻译：重复蛋白质的集合，每个重复蛋白质包含重复结构域，其包含一组连续重复模块，其中每个重复模块衍生自一个或多个重复单元，并且其中重复单元包含框架残基，其有助于重复单元的折叠拓扑或有助于与相邻重复单元的相互作用，以及有助于与靶物质相互作用的靶相互作用残基，其中所述重复蛋白与所述集合中的至少一个氨基酸中的其它重复蛋白不同描述重复模块的位置。

6. 发明申请

US20100081580A1 In vivo library-versus-library selection of optimized protein-protein interactions 审中-公开
标题翻译：体内文库对文库选择优化的蛋白质 - 蛋白质相互作用
公开(公告)号：US20100081580A1
公开(公告)日：2010-04-01
申请号：US12591731
申请日：2009-11-30
申请人： Stephen William Watson Michnick , Joelle N. Pelletier , Katja M. Arndt , Andreas Pluckthun
发明人： Stephen William Watson Michnick , Joelle N. Pelletier , Katja M. Arndt , Andreas Pluckthun
IPC分类号： C40B30/00
CPC分类号： C40B30/04 , C07K14/43595 , C07K2319/00 , C12N15/1055 , C12Q1/32 , G01N33/542 , G01N33/6845 , Y02A90/26
摘要： The present invention describes a rapid and efficient in vivo library-versus-library screening strategy for identifying optimally interacting pairs of heterodimerizing polypeptides. It allows for the screening of a protein library against a second protein library, rather than against a single bait protein, and thus has numerous applications in the study of protein-protein interactions. Additionally, it allows for the application of different selection stringencies. Two leucine zipper libraries, semi-randomized at the positions adjacent to the hydrophobic core, were genetically fused to either one of two designed fragments of the enzyme murine dihydrofolate reductase (mDHFR), and cotransformed into E. coli. Interaction between the library polypeptides was required for reconstitution of the enzymatic activity of mDHFR, allowing bacterial growth. Analysis of the resulting colonies revealed important biases in the zipper sequences relative to the original libraries, which are consistent with selection for stable, heterodimerizing pairs. Using more weakly associating mDHFR fragments, we increased the stringency of selection. We enriched the best performing leucine zipper pairs by multiple passaging of the pooled, selected colonies in liquid culture, as the best pairs allowed for better bacterial propagation. This competitive growth allowed small differences among the pairs to be amplified, and different sequence positions were enriched at different rates. We applied these selection processes to a library-versus-library sample of 2.0×106 combinations, and selected a novel leucine zipper pair which may be appropriate for use in further in vivo heterodimerization strategies.
摘要翻译：本发明描述了快速和有效的体内文库对文库筛选策略，用于鉴定异构二聚化多肽的最佳相互作用对。它允许针对第二蛋白质文库筛选蛋白质文库，而不是针对单个诱饵蛋白质，因此在蛋白质 - 蛋白质相互作用的研究中具有许多应用。此外，它允许应用不同的选择严格性。两个亮氨酸拉链文库在与疏水核心相邻的位置半随机化，与酶二氢叶酸还原酶（mDHFR）的两个设计的片段中的任一个遗传融合，并共转化到大肠杆菌中。需要文库多肽之间的相互作用来重构mDHFR的酶活性，从而允许细菌生长。所得菌落的分析揭示了相对于原始文库的拉链序列的重要偏倚，这与选择稳定的异源二聚体对一致。使用更弱的关联mDHFR片段，我们增加了选择的严格性。我们通过在液体培养中多次传代合并的选定菌落，丰富了表现最好的亮氨酸拉链对，因为最佳对允许更好的细菌繁殖。这种竞争性增长允许放大对中的小差异，并且以不同的速率富集不同的序列位置。我们将这些选择过程应用于2.0×106组合的库与文库样本，并选择了可能适用于进一步体内异二聚化策略的新型亮氨酸拉链对。

7. 发明申请

US20080026948A1 Protein (poly)peptides libraries 失效
标题翻译：蛋白质（多）肽文库
公开(公告)号：US20080026948A1
公开(公告)日：2008-01-31
申请号：US11642593
申请日：2006-12-21
申请人： Achim Knappik , Peter Pack , Liming Ge , Simon Moroney , Andreas Pluckthun
发明人： Achim Knappik , Peter Pack , Liming Ge , Simon Moroney , Andreas Pluckthun
IPC分类号： C40B20/06 , C07K16/00
CPC分类号： C07K1/047 , C07K16/00 , C07K16/005 , C07K16/18 , C07K16/242 , C07K16/26 , C07K16/2854 , C07K16/2896 , C07K16/44 , C07K2317/21 , C07K2317/565 , C07K2317/622 , C07K2319/00 , C12N15/10 , C12N15/1037 , C40B40/02
摘要： The present invention relates to synthetic DNA sequence which encode one or more collections of homologous proteins(poly)peptides, and methods for generating and applying libraries of these DNA sequences. In particular, the invention relates to the preparation of a library of human-derived antibody genes by the use of synthetic consensus sequences which cover the structural repertoire of antibodies encoded in the human genome. Furthermore, the invention relates to the use of a single consensus antibody gene as a universal framework for highly diverse antibody libraries.
摘要翻译：本发明涉及编码一个或多个同源蛋白（多）肽集合的合成DNA序列，以及用于产生和应用这些DNA序列的文库的方法。特别地，本发明涉及通过使用覆盖人类基因组中编码的抗体结构谱的合成共有序列来制备人源抗体基因文库。此外，本发明涉及单一共有抗体基因作为高度多样化抗体文库的通用框架的用途。

8. 发明申请

US20060159683A1 Novel method for the stabilization of chimeric immunoglobulins or immunoglobulin fragments, and stabilized anti-EGP-2 scFv fragment 有权
公开(公告)号：US20060159683A1
公开(公告)日：2006-07-20
申请号：US11024877
申请日：2004-12-30
申请人： Andreas Pluckthun , Annemarie Honegger , Jorg Willuda
发明人： Andreas Pluckthun , Annemarie Honegger , Jorg Willuda
IPC分类号： A61K39/395 , C07K16/30 , G01N33/574
CPC分类号： C07K16/00 , A61K47/6813 , A61K47/6819 , A61K47/6821 , A61K47/6823 , A61K47/6825 , A61K47/6829 , A61K47/6851 , A61K47/6877 , A61K51/1045 , A61K51/1087 , A61K2039/505 , C07K16/06 , C07K16/28 , C07K16/30 , C07K16/465 , C07K2317/565 , C07K2317/567 , C07K2317/622 , C07K2317/92
摘要： The present invention relates to a method for stabilizing chimeric immunoglobulins or immunoglobulin fragments. Furthermore, the invention also provides a stabilized anti-EGP-2 scFv fragment.

9. 发明授权

US09006389B2 Repeat protein from collection of repeat proteins comprising repeat modules 有权
标题翻译：从重复蛋白质的重复蛋白重复，包括重复模块
公开(公告)号：US09006389B2
公开(公告)日：2015-04-14
申请号：US13330336
申请日：2011-12-19
申请人： Michael Tobias Stumpp , Patrik Forrer , Hans Kaspar Binz , Andreas Pluckthun
发明人： Michael Tobias Stumpp , Patrik Forrer , Hans Kaspar Binz , Andreas Pluckthun
IPC分类号： A61K38/00 , C07K14/00 , A61K38/16 , C12N15/10 , C07K14/47
CPC分类号： C12N15/1055 , A61K38/00 , A61K38/16 , C07K14/00 , C07K14/47 , C07K14/4703 , C12N15/10 , C12N15/1037 , C12N15/1044 , C12N15/1093
摘要： A repeat protein from a collection of repeat proteins, wherein each repeat protein of said collection comprises a repeat domain, which comprises a set of consecutive repeat modules, wherein the repeat modules have the same fold and stack tightly to create a superhelical structure having a joint hydrophobic core, wherein each of the repeat modules is derived from one or more repeat units and wherein the repeat units comprise framework residues, which contribute to the folding topology of the repeat unit or contribute to an interaction with a neighboring repeat unit, and target interaction residues, which contribute to an interaction with a target substance, wherein the repeat proteins of the collection differ from other repeat proteins in the collection in at least one amino acid position of the repeat modules is described as are related pharmaceuticals and nucleic acid molecules.
摘要翻译：来自重复蛋白质的重复蛋白质，其中所述收集物的每个重复蛋白质包含重复结构域，其包含一组连续重复模块，其中所述重复模块具有相同的折叠并且紧密堆叠以产生具有关节的超螺旋结构疏水核心，其中每个重复模块衍生自一个或多个重复单元，并且其中重复单元包括框架残基，其有助于重复单元的折叠拓扑或有助于与相邻重复单元的相互作用，以及靶相互作用有助于与靶物质相互作用的残基，其中所述集合的重复蛋白质与所述重复模块的至少一个氨基酸位置中的所述集合中的其它重复蛋白质不同描述为相关药物和核酸分子。

10. 发明授权

US07625700B2 In vivo library-versus-library selection of optimized protein-protein interactions 失效
标题翻译：体内文库对文库选择优化的蛋白质 - 蛋白质相互作用
公开(公告)号：US07625700B2
公开(公告)日：2009-12-01
申请号：US11134253
申请日：2005-05-23
申请人： Stephen William Watson Michnick , Joelle N Pelletier , Katja M. Arndt , Andreas Pluckthun
发明人： Stephen William Watson Michnick , Joelle N Pelletier , Katja M. Arndt , Andreas Pluckthun
IPC分类号： C12Q1/68 , C12Q1/70 , C12P21/04 , C12N15/00 , C07H21/02 , C07H21/04
CPC分类号： C40B30/04 , C07K14/43595 , C07K2319/00 , C12N15/1055 , C12Q1/32 , G01N33/542 , G01N33/6845 , Y02A90/26
摘要： The present invention describes a rapid and efficient in vivo library-versus-library screening strategy for identifying optimally interacting pairs of heterodimerizing polypeptides. It allows for the screening of a protein library against a second protein library and therefore finds numerous applications in the study of protein-protein interactions. Two leucine zipper libraries, semi-randomized at the positions adjacent to the hydrophobic core, were genetically fused to either one of two designed fragments of the enzyme murine dihydrofolate reductase (mDHFR), and cotransformed into E. coli. Interaction between the library polypeptides was required for reconstitution of the enzymatic activity of mDHFR, allowing bacterial growth. Using more weakly associating mDHFR fragments, we increased the stringency of selection. We applied these selection processes to a library-versus-library sample of 2.0×106 combinations, and selected a novel leucine zipper pair which may be appropriate for use in further in vivo heterodimerization strategies.
摘要翻译：本发明描述了快速和有效的体内文库对文库筛选策略，用于鉴定异构二聚化多肽的最佳相互作用对。它允许对第二个蛋白质文库进行蛋白质文库的筛选，因此可以在研究蛋白质 - 蛋白质相互作用中发现许多应用。两个亮氨酸拉链文库在与疏水核心相邻的位置半随机化，与酶二氢叶酸还原酶（mDHFR）的两个设计的片段中的任一个遗传融合，并共转化到大肠杆菌中。需要文库多肽之间的相互作用来重构mDHFR的酶活性，从而允许细菌生长。使用更弱的关联mDHFR片段，我们增加了选择的严格性。我们将这些选择过程应用于2.0×10 6组合的文库与文库样品，并选择了可能适用于进一步体内异二聚化策略的新型亮氨酸拉链对。

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