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41. 发明授权

US09827280B2 Transgenic microalgae and use thereof for oral delivery of proteins 有权
公开(公告)号：US09827280B2
公开(公告)日：2017-11-28
申请号：US14628891
申请日：2015-02-23
申请人： Transalgae Israel Ltd.
发明人： Shiri Moshitzky , Doron Eisenstadt , Guy Levi , Ofra Chen
IPC分类号： A61K36/02 , C12N1/13 , A61K38/22 , C07K14/575 , A61K38/27 , C12N1/12 , C07K14/61 , C07K14/46 , A23K10/16 , A23K10/18 , A23K50/75 , A23K50/80
CPC分类号： A61K36/02 , A23K10/16 , A23K10/18 , A23K50/75 , A23K50/80 , A61K38/22 , A61K38/27 , C07K14/461 , C07K14/61 , C07K2319/04 , C07K2319/05 , C07K2319/06 , C12N1/12
摘要： Transgenic microalgae expressing at least one exogenous biologically active protein. The protein-expressing microalgae are used for the oral delivery of the biologically active protein to the target organism in its intact and functional form. The exogenous protein, expressed in algae, is characterized by being biologically active, exerting at least one specific activity having a beneficial effect on the subject consuming the algae. The transgenic microalgae are used as animal food for aquatic or land animals welfare or as food supplement for human healthcare.

42. 发明授权

US08932825B2 Method to engineer mammalian-type carbohydrate structures 有权
标题翻译：哺乳动物型碳水化合物结构的设计方法
公开(公告)号：US08932825B2
公开(公告)日：2015-01-13
申请号：US10500240
申请日：2002-12-24
申请人： Stefan Wildt , Robert Gordon Miele , Juergen Hermann Nett , Robert C. Davidson
发明人： Stefan Wildt , Robert Gordon Miele , Juergen Hermann Nett , Robert C. Davidson
IPC分类号： G01N33/569 , C12P1/02 , C12N1/00 , A61K51/00 , C12N9/10 , C12P21/00
CPC分类号： C12N15/815 , A01K2217/075 , C07K14/705 , C07K2319/05 , C12N9/1051 , C12N9/2402 , C12P21/005 , C12Y204/01143 , C12Y204/01144 , C12Y204/01145 , C12Y204/01155 , C12Y302/01024
摘要： The present invention relates to host cells having modified lipid-linked oligosaccharides which may be modified further by heterologous expression of a set of glycosyltransferases, sugar transporters and mannosidases to become host-strains for the production of mammalian, e.g., human therapeutic glycoproteins. The process provides an engineered host cell which can be used to express and target any desirable gene(s) involved in glycosylation. Host cells with modified lipid-linked oligosaccharides are created or selected. N-glycans made in the engineered host cells have a GlcNAcMan3GlcNAc2 core structure which may then be modified further by heterologous expression of one or more enzymes, e.g., glycosyl-transferases, sugar transporters and mannosidases, to yield human-like glycoproteins. For the production of therapeutic proteins, this method may be adapted to engineer cell lines in which any desired glycosylation structure may be obtained.
摘要翻译：本发明涉及具有经修饰的脂质连接的寡糖的宿主细胞，其可通过异源表达一组糖基转移酶，糖转运体和甘露糖苷酶进一步修饰，以成为用于产生哺乳动物例如人治疗性糖蛋白的宿主菌株。该方法提供了可用于表达和靶向参与糖基化的任何所需基因的工程化宿主细胞。制备或选择具有修饰的脂质连接寡糖的宿主细胞。在工程化的宿主细胞中制备的N-聚糖具有GlcNAcMan3GlcNAc2核心结构，然后可以通过异源表达一种或多种酶，例如糖基转移酶，糖转运蛋白和甘露糖苷酶来进一步修饰，以产生人样糖蛋白。为了生产治疗性蛋白质，该方法可适用于工程化可能获得任何所需糖基化结构的细胞系。

43. 发明申请

US20130018177A1 Production of Sialylated N-Glycans in Lower Eukaryotes 审中-公开
标题翻译：在低等真核生物中生产唾液酸化N-糖
公开(公告)号：US20130018177A1
公开(公告)日：2013-01-17
申请号：US13554126
申请日：2012-07-20
申请人： Stephen R. Hamilton
发明人： Stephen R. Hamilton
IPC分类号： C12P21/00 , C12N1/15 , C12N1/19 , C07K14/435 , C12N15/54
CPC分类号： C12P21/005 , C07K2319/036 , C07K2319/05 , C07K2319/055 , C12N9/1081 , C12N9/2402 , C12Y302/01018
摘要： The present invention relates to eukaryotic host cells which have been modified to produce sialylated glycoproteins by the heterologous expression of a set of glycosyltransferases, including sialyltransferase and/or trans-sialidase, to become host-strains for the production of mammalian, e.g., human therapeutic glycoproteins. Novel eukaryotic host cells expressing a CMP-sialic acid biosynthetic pathway for the production of sialylated glycoproteins are also provided. The invention provides nucleic acid molecules and combinatorial libraries which can be used to successfully target and express mammalian enzymatic activities (such as those involved in sialylation) to intracellular compartments in a eukaryotic host cell. The process provides an engineered host cell which can be used to express and target any desirable gene(s) involved in glycosylation.
摘要翻译：本发明涉及通过异源表达一组糖基转移酶（包括唾液酸转移酶和/或反式唾液酸酶）而被修饰以产生唾液酸化糖蛋白的真核宿主细胞，以成为用于产生哺乳动物例如人类治疗的宿主菌株糖蛋白。还提供了表达用于产生唾液酸化糖蛋白的CMP-唾液酸生物合成途径的新型真核宿主细胞。本发明提供核酸分子和组合文库，其可用于成功靶向并表达哺乳动物酶活性（例如涉及唾液酸化的那些）到真核宿主细胞中的细胞内区室。该方法提供了可用于表达和靶向参与糖基化的任何所需基因的工程化宿主细胞。

44. 发明授权

US08268609B2 Production of sialylated N-glycans in lower eukaryotes 失效
标题翻译：在低等真核生物中生产唾液酸化的N-聚糖
公开(公告)号：US08268609B2
公开(公告)日：2012-09-18
申请号：US12819305
申请日：2010-06-21
申请人： Stephen R. Hamilton
发明人： Stephen R. Hamilton
IPC分类号： C12N1/19
CPC分类号： C12P21/005 , C07K2319/036 , C07K2319/05 , C07K2319/055 , C12N9/1081 , C12N9/2402 , C12Y302/01018
摘要： The present invention relates to eukaryotic host cells which have been modified to produce sialylated glycoproteins by the heterologous expression of a set of glycosyltransferases, including sialyltransferase and/or trans-sialidase, to become host-strains for the production of mammalian, e.g., human therapeutic glycoproteins. Novel eukaryotic host cells expressing a CMP-sialic acid biosynthetic pathway for the production of sialylated glycoproteins are also provided. The invention provides nucleic acid molecules and combinatorial libraries which can be used to successfully target and express mammalian enzymatic activities (such as those involved in sialylation) to intracellular compartments in a eukaryotic host cell. The process provides an engineered host cell which can be used to express and target any desirable gene(s) involved in glycosylation.
摘要翻译：本发明涉及通过异源表达一组糖基转移酶（包括唾液酸转移酶和/或反式唾液酸酶）而被修饰以产生唾液酸化糖蛋白的真核宿主细胞，以成为用于产生哺乳动物例如人类治疗的宿主菌株糖蛋白。还提供了表达用于产生唾液酸化糖蛋白的CMP-唾液酸生物合成途径的新型真核宿主细胞。本发明提供核酸分子和组合文库，其可用于成功靶向并表达哺乳动物酶活性（例如涉及唾液酸化的那些）到真核宿主细胞中的细胞内区室。该方法提供了可用于表达和靶向参与糖基化的任何所需基因的工程化宿主细胞。

45. 发明授权

US08211691B2 Methods for producing modified glycoproteins 有权
标题翻译：生产改性糖蛋白的方法
公开(公告)号：US08211691B2
公开(公告)日：2012-07-03
申请号：US11981408
申请日：2007-10-30
申请人： Tillman U. Gerngross
发明人： Tillman U. Gerngross
IPC分类号： C12N15/04 , C12N1/16 , A61K38/00
CPC分类号： C12P21/005 , C07K2319/04 , C07K2319/05 , C12N1/14 , C12N9/1048 , C12N9/2488 , C12N15/79 , C12N15/80 , C12N15/81 , C12Y302/01 , C12Y302/01113
摘要： Cell lines having genetically modified glycosylation pathways that allow them to carry out a sequence of enzymatic reactions, which mimic the processing of glycoproteins in humans, have been developed. Recombinant proteins expressed in these engineered hosts yield glycoproteins more similar to their human counterparts. The lower eukaryotes, which ordinarily produce high-mannose containing N-glycans, including unicellular and multicellular fungi are modified to produce N-glycans such as Man5GlcNAc2 or other structures along human glycosylation pathways. This is achieved using a combination of engineering and/or selection of strains which: do not express certain enzymes which create the undesirable complex structures characteristic of the fungal glycoproteins, which express exogenous enzymes selected either to have optimal activity under the conditions present in the fungi where activity is desired, or which are targeted to an organelle where optimal activity is achieved, and combinations thereof wherein the genetically engineered eukaryote expresses multiple exogenous enzymes required to produce “human-like” glycoproteins.
摘要翻译：已经开发了具有遗传修饰的糖基化途径的细胞系，其允许它们进行模拟人类中糖蛋白的加工的酶反应序列。在这些工程化宿主中表达的重组蛋白产生更类似于人类对应物的糖蛋白。通常产生含有高甘露糖的N-聚糖（包括单细胞和多细胞真菌）的低等真核生物被修饰以产生N-聚糖如Man5GlcNAc2或沿着人糖基化途径的其它结构。这是使用以下工程和/或选择的组合实现的：不表达产生真菌糖蛋白特征的不合需要的复合结构的某些酶，其表达选择在真菌中存在的条件下具有最佳活性的外源酶其中需要活性或靶向实现最佳活性的细胞器，以及其组合，其中遗传工程真核生物表达产生“人样”糖蛋白所需的多种外源酶。

46. 发明授权

US07923430B2 Methods for producing modified glycoproteins 有权
标题翻译：生产改性糖蛋白的方法
公开(公告)号：US07923430B2
公开(公告)日：2011-04-12
申请号：US11249061
申请日：2005-10-11
申请人： Tillman U. Gerngross
发明人： Tillman U. Gerngross
IPC分类号： A61K31/715 , A61K31/7028 , A61K31/702 , A61K38/00
CPC分类号： C12P21/005 , C07K2319/04 , C07K2319/05 , C12N1/14 , C12N9/1048 , C12N9/2488 , C12N15/79 , C12N15/80 , C12N15/81 , C12Y302/01 , C12Y302/01113
摘要： Cell lines having genetically modified glycosylation pathways that allow them to carry out a sequence of enzymatic reactions, which mimic the processing of glycoproteins in humans, have been developed. The lower eukaryotes, which ordinarily produce high-mannose containing N-glycans, including unicellular and multicellular fungi are modified to produce N-glycans such as Man5GlcNAc2 or other structures along human glycosylation pathways. This is achieved using a combination of engineering and/or selection of strains which: do not express certain enzymes which create the undesirable complex structures characteristic of the fungal glycoproteins, which express exogenous enzymes selected either to have optimal activity under the conditions present in the fungi where activity is desired, or which are targeted to an organelle where optimal activity is achieved, and combinations thereof wherein the genetically engineered eukaryote expresses multiple exogenous enzymes required to produce “human-like” glycoproteins.
摘要翻译：已经开发了具有遗传修饰的糖基化途径的细胞系，其允许它们进行模拟人类中糖蛋白的加工的酶反应序列。通常产生含有高甘露糖的N-聚糖（包括单细胞和多细胞真菌）的低等真核生物被修饰以产生N-聚糖如Man5GlcNAc2或沿着人糖基化途径的其它结构。这是使用以下工程和/或选择的组合实现的：不表达产生真菌糖蛋白特征的不合需要的复合结构的某些酶，其表达选择在真菌中存在的条件下具有最佳活性的外源酶其中需要活性或靶向实现最佳活性的细胞器，以及其组合，其中遗传工程真核生物表达产生“人样”糖蛋白所需的多种外源酶。

47. 发明授权

US07629163B2 Methods for producing modified glycoproteins 失效
标题翻译：生产改性糖蛋白的方法
公开(公告)号：US07629163B2
公开(公告)日：2009-12-08
申请号：US11271235
申请日：2005-11-10
申请人： Tillman U. Gerngross
发明人： Tillman U. Gerngross
IPC分类号： C12N1/15 , C07K14/00
CPC分类号： C12P21/005 , C07K2319/04 , C07K2319/05 , C12N1/14 , C12N9/1048 , C12N9/2488 , C12N15/79 , C12N15/80 , C12N15/81 , C12Y302/01 , C12Y302/01113
摘要： Cell lines having genetically modified glycosylation pathways that allow them to carry out a sequence of enzymatic reactions, which mimic the processing of glycoproteins in humans, have been developed. Recombinant proteins expressed in these engineered hosts yield glycoproteins more similar, if not substantially identical, to their human counterparts. The lower eukaryotes, which ordinarily produce high-mannose containing N-glycans, including unicellular and multicellular fungi are modified to produce N-glycans such as Man5GlcNAc2 or other structures along human glycosylation pathways. This is achieved using a combination of engineering and/or selection of strains which: do not express certain enzymes which create the undesirable complex structures characteristic of the fungal glycoproteins, which express exogenous enzymes selected either to have optimal activity under the conditions present in the fungi where activity is desired, or which are targeted to an organelle where optimal activity is achieved, and combinations thereof wherein the genetically engineered eukaryote expresses multiple exogenous enzymes required to produce “human-like” glycoproteins.
摘要翻译：已经开发了具有遗传修饰的糖基化途径的细胞系，其允许它们进行模拟人类中糖蛋白的加工的酶反应序列。在这些工程化宿主中表达的重组蛋白可以产生与其对应物更相似的糖蛋白（如果基本上不相同）。通常产生含有高甘露糖的N-聚糖（包括单细胞和多细胞真菌）的低等真核生物被修饰以产生N-聚糖如Man5GlcNAc2或沿着人糖基化途径的其它结构。这是使用以下工程和/或选择的组合实现的：不表达产生真菌糖蛋白特征的不合需要的复合结构的某些酶，其表达选择在真菌中存在的条件下具有最佳活性的外源酶其中需要活性或靶向实现最佳活性的细胞器，以及其组合，其中遗传工程真核生物表达产生“人样”糖蛋白所需的多种外源酶。

48. 发明授权

US07326681B2 Methods for producing modified glycoproteins 有权
标题翻译：生产改性糖蛋白的方法
公开(公告)号：US07326681B2
公开(公告)日：2008-02-05
申请号：US11240432
申请日：2005-09-30
申请人： Tillman U. Gerngross
发明人： Tillman U. Gerngross
IPC分类号： A61K38/14 , A61K38/00
CPC分类号： C12P21/005 , C07K2319/04 , C07K2319/05 , C12N1/14 , C12N9/1048 , C12N9/2488 , C12N15/79 , C12N15/80 , C12N15/81 , C12Y302/01 , C12Y302/01113
摘要： Cell lines having genetically modified glycosylation pathways that allow them to carry out a sequence of enzymatic reactions, which mimic the processing of glycoproteins in humans, have been developed. Recombinant proteins expressed in these engineered hosts yield glycoproteins more similar, if not substantially identical, to their human counterparts. The lower eukaryotes, which ordinarily produce high-mannose containing N-glycans, including unicellular and multicellular fungi are modified to produce N-glycans such as Man5GlcNAc2 or other structures along human glycosylation pathways. This is achieved using a combination of engineering and/or selection of strains which: do not express certain enzymes which create the undesirable complex structures characteristic of the fungal giycoproteins, which express exogenous enzymes selected either to have optimal activity under the conditions present in the fungi where activity is desired, or which are targeted to an organelle where optimal activity is achieved, and combinations thereof wherein the genetically engineered eukaryote expresses multiple exogenous enzymes required to produce “human-like” glycoproteins.
摘要翻译：已经开发了具有遗传修饰的糖基化途径的细胞系，其允许它们进行模拟人类中糖蛋白的加工的酶反应序列。在这些工程化宿主中表达的重组蛋白可以产生与其对应物更相似的糖蛋白（如果基本上不相同）。通常产生含有高甘露糖的N-聚糖（包括单细胞和多细胞真菌）的低等真核生物被修饰以产生N-聚糖，例如Man 3或GlcNAc 2 N或其它结构沿着人类糖基化途径。这是使用以下工程和/或选择的组合实现的：不表达产生真菌糖蛋白特征的不合需要的复合结构的某些酶，其表达选择在真菌中存在的条件下具有最佳活性的外源酶其中需要活性或靶向实现最佳活性的细胞器，以及其组合，其中遗传工程真核生物表达产生“人样”糖蛋白所需的多种外源酶。

49. 发明申请

US20060148035A1 Methods for producing modified glycoproteins 失效
标题翻译：生产改性糖蛋白的方法
公开(公告)号：US20060148035A1
公开(公告)日：2006-07-06
申请号：US11271235
申请日：2005-11-10
申请人： Tillman Gerngross
发明人： Tillman Gerngross
IPC分类号： C12P21/06 , C07H21/04 , C12N9/24 , C12N1/18 , C12N15/74
CPC分类号： C12P21/005 , C07K2319/04 , C07K2319/05 , C12N1/14 , C12N9/1048 , C12N9/2488 , C12N15/79 , C12N15/80 , C12N15/81 , C12Y302/01 , C12Y302/01113
摘要： Cell lines having genetically modified glycosylation pathways that allow them to carry out a sequence of enzymatic reactions, which mimic the processing of glycoproteins in humans, have been developed. Recombinant proteins expressed in these engineered hosts yield glycoproteins more similar, if not substantially identical, to their human counterparts. The lower eukaryotes, which ordinarily produce high-mannose containing N-glycans, including unicellular and multicellular fungi are modified to produce N-glycans such as Man5GlcNAc2 or other structures along human glycosylation pathways. This is achieved using a combination of engineering and/or selection of strains which: do not express certain enzymes which create the undesirable complex structures characteristic of the fungal glycoproteins, which express exogenous enzymes selected either to have optimal activity under the conditions present in the fungi where activity is desired, or which are targeted to an organelle where optimal activity is achieved, and combinations thereof wherein the genetically engineered eukaryote expresses multiple exogenous enzymes required to produce “human-like” glycoproteins.
摘要翻译：已经开发了具有遗传修饰的糖基化途径的细胞系，其允许它们进行模拟人类中糖蛋白的加工的酶反应序列。在这些工程化宿主中表达的重组蛋白可以产生与其对应物更相似的糖蛋白（如果基本上不相同）。通常产生含有高甘露糖的N-聚糖（包括单细胞和多细胞真菌）的低等真核生物被修饰以产生N-聚糖，例如Man 3或GlcNAc 2 N或其它结构沿着人类糖基化途径。这是使用以下工程和/或选择的组合实现的：不表达产生真菌糖蛋白特征的不合需要的复合结构的某些酶，其表达选择在真菌中存在的条件下具有最佳活性的外源酶其中需要活性或靶向实现最佳活性的细胞器，以及其组合，其中遗传工程真核生物表达产生“人样”糖蛋白所需的多种外源酶。

50. 发明申请

US20050208617A1 N-acetylglucosamintransferase III expression in lower eukaryotes 失效
标题翻译： N-乙酰葡萄糖转移酶III在低等真核生物中的表达
公开(公告)号：US20050208617A1
公开(公告)日：2005-09-22
申请号：US10680963
申请日：2003-10-07
申请人： Piotr Bobrowicz , Stephen Hamilton , Tilman Gerngross , Stefan Wildt , Byung-Kwon Choi , Juergen Nett , Robert Davidson
发明人： Piotr Bobrowicz , Stephen Hamilton , Tilman Gerngross , Stefan Wildt , Byung-Kwon Choi , Juergen Nett , Robert Davidson
IPC分类号： A61K38/00 , C07K14/39 , C07K14/47 , C12N1/18 , C12N9/10 , C12N15/12 , C12N15/74 , C12P21/00 , C12P21/06
CPC分类号： C07K14/39 , A61K38/00 , C07K2319/05 , C12N9/1051 , C12P21/005 , Y02A50/396
摘要： The present invention relates to eukaryotic host cells having modified oligosaccharides which may be modified further by heterologous expression of a set of glycosyltransferases, sugar transporters and mannosidases to become host-strains for the production of mammalian, e.g., human therapeutic glycoproteins. The process provides an engineered host cell which can be used to express and target any desirable gene(s) involved in glycosylation. Host cells with modified lipid-linked oligosaccharides are created or selected. N-glycans made in the engineered host cells exhibit GnTIII activity, which produce bisected N-glycan structures and may be modified further by heterologous expression of one or more enzymes, e.g., glycosyltransferases, sugar transporters and mannosidases, to yield human-like glycoproteins. For the production of therapeutic proteins, this method may be adapted to engineer cell lines in which any desired glycosylation structure may be obtained.
摘要翻译：本发明涉及具有修饰的寡糖的真核宿主细胞，其可以通过异源表达一组糖基转移酶，糖转运体和甘露糖苷酶进一步修饰，以成为用于产生哺乳动物例如人治疗性糖蛋白的宿主菌株。该方法提供了可用于表达和靶向参与糖基化的任何所需基因的工程化宿主细胞。制备或选择具有修饰的脂质连接寡糖的宿主细胞。在工程化的宿主细胞中制备的N-聚糖表现出GnTIII活性，其产生二等分的N-聚糖结构，并且可以通过异源表达一种或多种酶（例如糖基转移酶，糖转运蛋白和甘露糖苷酶）进一步修饰，以产生人样糖蛋白。为了生产治疗性蛋白质，该方法可适用于工程化可能获得任何所需糖基化结构的细胞系。

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