专利快速检索-快速检索全球专利，免费商用专利数据库-IPRDB

1. 发明授权

US07449316B2 UDP-galactose: β-D-galactose-R 4-α-D-galactosyltransferase, α4Gal-T1 失效
标题翻译： UDP-半乳糖：β-D-半乳糖-R 4-α-D-半乳糖基转移酶，α4Gal-T1
公开(公告)号：US07449316B2
公开(公告)日：2008-11-11
申请号：US11317196
申请日：2005-12-22
申请人： Henrik Clausen , Eric Paul Bennett , Rudi Steffensen
发明人： Henrik Clausen , Eric Paul Bennett , Rudi Steffensen
IPC分类号： C12N9/10 , C12P19/00 , C12P21/04 , C12Q1/00 , C12Q1/68 , C12N1/20 , C12N15/00 , C07H21/04
CPC分类号： C12N9/1051
摘要： A novel gene defining a novel enzyme UDP-galactose: β-D-galactose-R 4-α-D-galactosyltransferase, termed α4Gal-T1, with unique enzymatic properties is disclosed. The invention provides isolated DNA molecules and DNA constructs encoding α4Gal-T1 and derivatives thereof by way of amino acid deletion, substitution or insertion exhibiting α4Gal-T1 activity, as well as cloning and expression vectors including such DNA, host cells comprising DNA encoding α4Gal-T1, and recombinant methods for providing α4Gal-T1. The enzyme α4Gal-T1 and α4Gal-active derivatives thereof are disclosed. Further, the invention discloses methods of obtaining α1, 4galactosyl glycosylated glycosphingolipids by use of an enzymatically active α4Gal-T1 protein thereof or by using cells stably transfected with a vector including DNA encoding an enzymatically active α4Gal-T1 protein as an expression system for recombinant production of such glycosphingolipids. Also a method for the identification of DNA sequence variations in the α4Gal-T1-coding exon by PCR, and detecting the presence of DNA sequence variation, are disclosed.
摘要翻译：公开了一种新颖的基因，其定义了具有独特的酶学性质的新型酶UDP-半乳糖：β-D-半乳糖-R 4-α-D-半乳糖基转移酶，称为α4Gal-T1。本发明提供分离的DNA分子和编码α4Gal-T1及其衍生物的DNA构建体，其通过氨基酸缺失，取代或插入表现出α4Gal-T1活性，以及克隆和表达载体，包括此类DNA，宿主细胞包含编码α4Gal- T1和用于提供α4Gal-T1的重组方法。公开了其α4Gal-T1和α4Gal活性衍生物。此外，本发明公开了通过使用其酶促活化的α4Gal-T1蛋白或通过使用包含编码酶活性α4Gal-T1蛋白的DNA稳定转染的细胞作为重组生产的表达系统来获得α1，4半乳糖基糖基化糖苷脂的方法的这种糖鞘醇。还公开了通过PCR鉴定α4Gal-T1编码外显子中的DNA序列变异以及检测DNA序列变异的存在的方法。

2. 发明授权

US07670816B2 UDP-galactose:β-D-galactose-R 4-α-D-galactosyltransferase, α4gal-T1 失效
标题翻译： UDP-半乳糖：β-半乳糖-R 4-α-D-半乳糖基转移酶，α4gal-T1
公开(公告)号：US07670816B2
公开(公告)日：2010-03-02
申请号：US12244921
申请日：2008-10-03
申请人： Henrik Clausen , Rudi Steffensen , Eric Paul Bennett
发明人： Henrik Clausen , Rudi Steffensen , Eric Paul Bennett
IPC分类号： C12N9/10 , C12P19/00 , C12P21/04 , C12Q1/00 , C12Q1/68 , C12N1/20 , C12N15/00 , C07H21/04
CPC分类号： C12N9/1051
摘要： A novel gene defining a novel enzyme UDP-galactose: β-D-galactose-R 4-α-D-galactosyltransferase, termed α4Gal-T1, with unique enzymatic properties is disclosed. The invention provides isolated DNA molecules and DNA constructs encoding α4Gal-T1 and derivatives thereof by way of amino acid deletion, substitution or insertion exhibiting α4Gal-T1 activity, as well as cloning and expression vectors including such DNA, host cells comprising DNA encoding α4Gal-T1, and recombinant methods for providing α4Gal-T1. The enzyme α4Gal-T1 and α4Gal-active derivatives thereof are disclosed. Further, the invention discloses methods of obtaining α1, 4galactosyl glycosylated glycosphingolipids by use of an enzymatically active α4Gal-T1 protein thereof or by using cells stably transfected with a vector including DNA encoding an enzymatically active α4Gal-T1 protein as an expression system for recombinant production of such glycosphingolipids. Also a method for the identification of DNA sequence variations in the α4Gal-T1-coding exon by PCR, and detecting the presence of DNA sequence variation, are disclosed.
摘要翻译：公开了一种新颖的基因，其定义了具有独特的酶学性质的新型酶UDP-半乳糖：＆bgr; -D-半乳糖-R 4-α-D-半乳糖基转移酶，称为α4Gal-T1。本发明通过具有α4Gal-T1活性的氨基酸缺失，取代或插入以及包括这种DNA的克隆和表达载体，提供分离的DNA分子和编码α4Gal-T1及其衍生物的DNA构建体，宿主细胞包含编码α4Gal- T1和提供α4Gal-T1的重组方法。公开了酶α4Gal-T1和α4Gal-活性衍生物。此外，本发明公开了通过使用其酶促活性α4Gal-T1蛋白质或通过使用包含编码酶活性α4Gal-T1蛋白质的DNA的载体稳定转染的细胞作为重组生产的表达系统，获得α1，4半乳糖基糖基化糖苷脂的方法的这种糖鞘醇。还公开了通过PCR鉴定α4Gal-T1编码外显子中的DNA序列变异并检测DNA序列变异的存在的方法。

3. 发明授权

US07115404B2 UDP-galactose: β-D-galactose-R 4-α-D-galac-tosyltransferase, α4Gal-T1 有权
标题翻译： UDP-半乳糖：β-D-半乳糖-R 4-α-D-加酰甘油甲苯磺酰转移酶，α4Gal-T1
公开(公告)号：US07115404B2
公开(公告)日：2006-10-03
申请号：US10217335
申请日：2002-08-09
申请人： Henrik Clausen , Rudi Steffensen , Eric Paul Bennett
发明人： Henrik Clausen , Rudi Steffensen , Eric Paul Bennett
IPC分类号： C12N9/10 , C12N9/00 , C12P21/06 , C07H21/04
CPC分类号： C12N9/1051
摘要： A novel gene defining a novel enzyme UDP-Galactose: β-D-Galactose-R 4-α-D-galactosyltransferase, termed α4Gal-T1, with unique enzymatic properties is disclosed. The invention discloses isolated DNA molecules and DNA constructs encoding α4Gal-T1 and derivatives thereof by way of amino acid deletion, substitution or insertion exhibiting α4Gal-T1 activity, as well as cloning and expression vectors including such DNA, cells transfected with vectors, and recombinant methods for providing α4Gal-T1. The enzyme α4Gal-T1 and α4Gal-active derivatives thereof are disclosed. Further, the invention discloses methods of obtaining α1, 4galactosyl glycosylated glycosphingolipids by use of an enzymatically active α4Gal-T1 protein thereof or by using cells stably transfected with a vector including DNA encoding an enzymatically active α4Gal-T1 protein as an expression system for recombinant production of such glycosphingolipids. Also a method for the identification of DNA sequence variations in the α4Gal-T1-coding exon by PCR, and detecting the presence of DNA sequence variation, are disclosed.
摘要翻译：公开了一种新颖的基因，其定义了具有独特的酶学性质的新型酶UDP-Galactose：β-D-半乳糖-R 4-α-D-半乳糖基转移酶，称为α4Gal-T1。本发明公开了分离的DNA分子和编码α4Gal-T1及其衍生物的DNA构建体，其通过氨基酸缺失，取代或插入表现出α4Gal-T1活性，以及克隆和表达载体，包括此类DNA，用载体转染的细胞和重组提供α4Gal-T1的方法。公开了其α4Gal-T1和α4Gal活性衍生物。此外，本发明公开了通过使用其酶促活化的α4Gal-T1蛋白或通过使用包含编码酶活性α4Gal-T1蛋白的DNA稳定转染的细胞作为重组生产的表达系统来获得α1，4半乳糖基糖基化糖苷脂的方法的这种糖鞘醇。还公开了通过PCR鉴定α4Gal-T1编码外显子中的DNA序列变异并检测DNA序列变异的存在的方法。

4. 发明授权

US06558934B1 UDP-galactose: &bgr;-N-acetyl-glucosamine &bgr;-1,4-galactosyl-transferase, &bgr;4Gal-T2 失效
标题翻译： UDP-半乳糖：β-N-乙酰葡糖胺β-1,4-半乳糖转移酶，β4Gal-T2
公开(公告)号：US06558934B1
公开(公告)日：2003-05-06
申请号：US09118464
申请日：1998-07-17
申请人： Henrik Clausen , Eric Paul Bennett
发明人： Henrik Clausen , Eric Paul Bennett
IPC分类号： C12P2106
CPC分类号： C12N9/1051
摘要： A novel gene defining a novel enzyme in the UDP-D-galactose: &bgr;-N-acetyl-glucosamine &bgr;-1,4-galactosyltransferase family, termed &bgr;4Gal-T2, with unique enzymatic properties is disclosed. The enzymatic activity of &bgr;4Gal-T2 is shown to be distinct from that of previously identified enzymes of this gene family. The invention discloses isolated DNA molecules and DNA constructs encoding &bgr;4Gal-T2 and derivatives thereof by way of amino acid deletion, substitution or insertion exhibiting &bgr;4Gal-T2 activity, as well as cloning and expression vectors including such DNA, cells transfected with the vectors, and recombinant methods for providing &bgr;4Gal-T2. The enzyme &bgr;4Gal-T2 and &bgr;4Gal-T2-active derivatives thereof are disclosed, in particular soluble derivatives comprising the catalytically active domain of &bgr;4Gal-T2. Further, the invention discloses methods of obtaining &bgr;-1,4-galactosyl glycosylated saccharides, glycopeptides or glycoproteins by use of an enzymically active &bgr;4Gal-T2 protein or fusion protein thereof or by using cells stably transfected with a vector including DNA encoding an enzymatically active &bgr;4Gal-T2 protein as an expression system for recombinant production of such glycopeptides or glycoproteins. Also a method for the identification of DNA sequence variations in the &bgr;4Gal-T2 gene by isolating DNA from a patient, amplifying &bgr;4Gal-T2-coding exons by PCR, and detecting the presence of DNA sequence variation, are disclosed.
摘要翻译：公开了在UDP-D-半乳糖:β-N-乙酰葡糖胺β-1,4-半乳糖基转移酶家族中定义新的酶的新基因，称为β4Gal-T2，具有独特的酶学性质。 β4Gal-T2的酶活性显示与以前鉴定的该基因家族酶的活性不同。本发明公开了分离的DNA分子和编码β4Gal-T2的DNA构建体及其衍生物，通过氨基酸缺失，取代或插入显示β4Gal-T2活性，以及克隆和表达载体，包括这种DNA，用载体转染的细胞，以及提供β4Gal-T2的重组方法。公开了酶β4Gal-T2和β4Gal-T2-活性衍生物，特别是包含β4Gal-T2的催化活性结构域的可溶性衍生物。此外，本发明公开了通过使用酶活性β4Gal-T2蛋白或其融合蛋白或通过使用包含编码酶活性的DNA的载体稳定转染的细胞获得β-1,4-半乳糖基糖基化糖，糖肽或糖蛋白的方法 β4Gal-T2蛋白作为重组生产这种糖肽或糖蛋白的表达系统。还公开了通过从患者中分离DNA，通过PCR扩增β4Gal-T2编码外显子并检测DNA序列变异的存在来鉴定β4Gal-T2基因中的DNA序列变异的方法。

5. 发明授权

US5871990A UDP-N-acetyl-.alpha.-D-galactosamine: polypeptide N-acetylgalactosaminyltransferase, gAlnAc-T3 失效
标题翻译： UDP-N-乙酰基-α-D-半乳糖胺：多肽N-乙酰半乳糖胺基转移酶，gAlnAc-T3
公开(公告)号：US5871990A
公开(公告)日：1999-02-16
申请号：US648298
申请日：1996-05-15
申请人： Henrik Clausen , Eric Paul Bennett
发明人： Henrik Clausen , Eric Paul Bennett
IPC分类号： A61K39/00 , C12N9/10 , C07H21/04 , C12N1/20 , C12P21/06
CPC分类号： C12N9/1051 , A61K39/00
摘要： A novel gene defining a novel enzyme in the UDP-N-acetyl-.alpha.-D-galactosamine: polypeptide N-acetylgalactosaminyltransferase family, termed GalNAc-T3, with unique enzymatic properties is disclosed. The enzymatic activity of GalNAc-T3 is shown to be distinct from that of two previously identified enzymes of this gene family. The invention discloses isolated DNA molecules and DNA constructs encoding GalNAc-T3 and derivatives thereof by way of amino acid deletion, substitution or insertion exhibiting GalNAc-T3 activity, as well as cloning and expression vectors including such DNA, cells transfected with the vectors, and recombinant methods for providing GalNAc-T3. The enzyme GalNAc-T3 and GalNAc-T3-active derivatives thereof are disclosed, in particular soluble derivatives comprising the catalytically active domain of GalNAc-T3. Further, the invention discloses methods of obtaining N-acetylgalactosamine glycosylated peptides or proteins by use of an enzymically active GalNAc-T3 protein or fusion protein thereof or by using cells stably transfected with a vector including DNA encoding an enzymatically active GalNAc-T3 protein as an expression system for recombinant production of such glycopeptides or glycoproteins. Also a method for the production of a vaccine by modifying the O-glycosylation pattern of an eukaryotic cell, and a method for the identification of DNA sequence variations in the GalNAc-T3 gene by isolating DNA from a patient, amplifying GalNAc-T3-coding exons by PCR, and detecting the presence of DNA sequence variation, are disclosed.
摘要翻译：公开了一种在UDP-N-乙酰-α-D-半乳糖胺中定义新型酶的新基因：多肽N-乙酰半乳糖胺基转移酶家族，称为GalNAc-T3，具有独特的酶学性质。 GalNAc-T3的酶活性显示与该基因家族的两种先前鉴定的酶的酶活性不同。本发明公开了通过氨基酸缺失，取代或插入显示GalNAc-T3活性的编码GalNAc-T3及其衍生物的分离的DNA分子和DNA构建体，以及包含这种DNA的克隆和表达载体，用载体转染的细胞，以及提供GalNAc-T3的重组方法。公开了GalNAc-T3及其GalNAc-T3-活性衍生物，特别是包含GalNAc-T3的催化活性结构域的可溶性衍生物。此外，本发明公开了通过使用酶活性GalNAc-T3蛋白或其融合蛋白或通过使用包含编码酶活性GalNAc-T3蛋白的DNA的载体稳定转染的细胞来获得N-乙酰半乳糖胺糖基化肽或蛋白质的方法，用于重组生产这种糖肽或糖蛋白的表达系统。另外，通过改变真核细胞的O-糖基化模式来生产疫苗的方法，以及通过从患者中分离DNA来鉴定GalNAc-T3基因中的DNA序列变异的方法，扩增GalNAc-T3编码公开了通过PCR的外显子和检测DNA序列变异的存在。

6. 发明申请

US20080206810A1 Truncated St6galnaci Polypeptides and Nucleic Acids 审中-公开
标题翻译：截短的St6galnaci多肽和核酸
公开(公告)号：US20080206810A1
公开(公告)日：2008-08-28
申请号：US11597258
申请日：2005-06-03
申请人： Karl F. Johnson , David James Hakes , Ge Wei , Li Liu , Sami Saribas , Eric R. Sjoberg , Henrik Clausen , Eric Paul Bennett , Aliakbar Mobasseri
发明人： Karl F. Johnson , David James Hakes , Ge Wei , Li Liu , Sami Saribas , Eric R. Sjoberg , Henrik Clausen , Eric Paul Bennett , Aliakbar Mobasseri
IPC分类号： C12P21/04 , C12N9/10 , C12N15/11 , C12N15/00 , C12N5/06 , C12P19/26 , C12N5/04 , C12N1/20 , C12N1/00
CPC分类号： C12N9/1081 , C07K2319/20 , C07K2319/40
摘要： The present invention features compositions and methods related to truncated mutants of ST6GalNAcI. In particular, the invention features truncated human, mouse, and chicken ST6GalNAcI polypeptides. The invention also features nucleic acids encoding such truncated polypeptides, as well as vectors, host cells, expression systems, and methods of expressing and using such polypeptides.
摘要翻译：本发明涉及与ST6GalNAcI的截短突变体有关的组合物和方法。特别地，本发明的特征在于截短的人，小鼠和鸡ST6GalNAcI多肽。本发明还包括编码这种截短的多肽的核酸，以及载体，宿主细胞，表达系统以及表达和使用这些多肽的方法。

7. 发明授权

US07374919B2 UDP-galactose: β-N-acetyl-glucosamine β-1,4-galactosyltransferase, β4Gal-T2 有权
标题翻译： UDP-半乳糖：β-N-乙酰葡糖胺β-1,4-半乳糖基转移酶，β4Gal-T2
公开(公告)号：US07374919B2
公开(公告)日：2008-05-20
申请号：US11105796
申请日：2005-04-13
申请人： Henrik Clausen , Eric Paul Bennett
发明人： Henrik Clausen , Eric Paul Bennett
IPC分类号： C12N9/10 , C07H21/04
CPC分类号： C12N9/1051
摘要： A novel gene defining a novel enzyme in the UDP-D-galactose: b-N-acetyl-glucosamine β-1,4-galactosyltransferase family, termed β4Gal-T2, with unique enzymatic properties is disclosed. The enzymatic activity of β4Gal-T2 is shown to be distinct from that of previously identified enzymes of this gene family. The invention discloses isolated DNA molecules and DNA constructs encoding β4Gal-T2 and derivatives thereof by way of amino acid deletion, substitution or insertion exhibiting β4Gal-T2 activity, as well as cloning and expression vectors including such DNA, cells transfected with the vectors, and recombinant methods for providing β4Gal-T2. The enzyme β4Gal-T2 and β4Gal-T2-active derivatives thereof are disclosed, in particular soluble derivatives comprising the catalytically active domain of β4Gal-T2. Further, the invention discloses methods of obtaining β-1,4-galactosyl glycosylated saccharides, glycopeptides or glycoproteins by use of an enzymically active β4Gal-T2 protein or fusion protein thereof or by using cells stably transfected with a vector including DNA encoding an enzymatically active β4Gal-T2 protein as an expression system for recombinant production of such glycopeptides or glycoproteins. Also a method for the identification of DNA sequence variations in the β4Gal-T2 gene by isolating DNA from a patient, amplifying β4Gal-T2-coding exons by PCR, and detecting the presence of DNA sequence variation, are disclosed.
摘要翻译：公开了一种在UDP-D-半乳糖：b-N-乙酰葡糖胺β-1,4-半乳糖基转移酶家族中定义新的酶的新基因，称为β4Gal-T2，具有独特的酶学性质。 β4Gal-T2的酶活性显示与以前鉴定的该基因家族酶的活性不同。本发明公开了分离的DNA分子和编码β4Gal-T2的DNA构建体及其衍生物，通过氨基酸缺失，取代或插入显示β4Gal-T2活性，以及克隆和表达载体，包括这种DNA，用载体转染的细胞，以及提供β4Gal-T2的重组方法。公开了酶β4Gal-T2和β4Gal-T2-活性衍生物，特别是包含β4Gal-T2的催化活性结构域的可溶性衍生物。此外，本发明公开了通过使用酶活性β4Gal-T2蛋白或其融合蛋白或通过使用包含编码酶活性的DNA的载体稳定转染的细胞获得β-1,4-半乳糖基糖基化糖，糖肽或糖蛋白的方法 β4Gal-T2蛋白作为重组生产这种糖肽或糖蛋白的表达系统。还公开了通过从患者中分离DNA，通过PCR扩增β4Gal-T2编码外显子并检测DNA序列变异的存在来鉴定β4Gal-T2基因中的DNA序列变异的方法。

8. 发明授权

US07338932B2 Methods of modulating functions of polypeptide GalNAc-transferases and of screening test substances to find agents herefor, pharmaceutical compositions comprising such agents and the use of such agents for preparing medicaments 有权
标题翻译：调节多肽GalNAc转移酶的功能和筛选测试物质以找到本发明的药剂的方法，包含这些药剂的药物组合物以及这些药物用于制备药物的用途
公开(公告)号：US07338932B2
公开(公告)日：2008-03-04
申请号：US10292896
申请日：2002-11-12
申请人： Henrik Clausen , Eric Paul Bennett , Helle Hassan , Celso Albuquerque Reis
发明人： Henrik Clausen , Eric Paul Bennett , Helle Hassan , Celso Albuquerque Reis
IPC分类号： A61K38/16 , A61K38/00 , C07H1/00 , C07H5/04 , C07H19/00
CPC分类号： A61K38/14 , A61K38/1709 , C12N9/1051 , C12Q1/48 , G01N2500/00 , G01N2500/10
摘要： Attachment of O-glycans to proteins is controlled by a large family of homologous polypeptide GalNAc-transferases. Polypeptide GalNAc-transferases contain a C-terminal sequence with similarity to lectins. This invention discloses that the putative lectin domains of GalNAc-transferase isoforms, GalNAc-T4, -T7, -T2, and -T3, are functional and recognize carbohydrates, glycopeptides, and peptides and discloses the lectin domains of GalNAc-T1-T16. These lectin domains have different binding specificities and modulate the functions of GalNAc-transferase isoforms differently. Novel methods for identification of inhibitors or modulators of binding activities mediated by lectin domains of polypeptide GalNAc-transferases are disclosed. Direct binding activity of GalNAc-transferase lectins has been demonstrated for the first time and methods to measure lectin mediated binding of isolated lectins or enzymes with lectin domains are disclosed. The present invention specifically discloses a novel selective inhibitor of polypeptide GalNAc-transferase lectin domains, which provides a major advancement in that this inhibitor and related inhibitors sharing common characteristics of activity bind lectin domains without serving as acceptor substrate for glycosyltransferases involved in synthesis of O-glycans. This inhibitor is represented by the β-anomeric configuration of GalNAc-benzyl, GalNAcβ-benzyl. Methods for inhibiting intracellular transport, cell surface expression, and secretion of mucins and O-glycosylated glycoproteins without affecting O-glycosylation processing are disclosed using the novel selective inhibitor identified.
摘要翻译： O-聚糖对蛋白质的附着由大量的同源多肽GalNAc转移酶家族控制。多肽GalNAc转移酶含有与凝集素相似的C-末端序列。本发明公开了GalNAc转移酶同工型，GalNAc-T4，-T7，-T2和-T3的推定凝集素结构域是功能性的并识别碳水化合物，糖肽和肽，并公开了GalNAc-T1-T16的凝集素结构域。这些凝集素结构域具有不同的结合特异性，并且不同地调节GalNAc转移酶同工型的功能。公开了用于鉴定由多肽GalNAc转移酶的凝集素结构域介导的结合活性的抑制剂或调节剂的新方法。已经首次证明了GalNAc转移酶凝集素的直接结合活性，并且公开了测定凝血素介导的分离的凝集素或酶与凝集素结构域的结合的方法。本发明特别公开了一种新型多肽GalNAc转移酶凝集素结构域的选择性抑制剂，其提供了主要进展，即该抑制剂和相关抑制剂共享活性的共同特征结合凝集素结构域，而不用作参与合成O-糖蛋白的糖基转移酶的受体底物，聚糖。该抑制剂由GalNAc-苄基，GalNAcβ-苄基的β-异头构型表示。使用鉴定的新型选择性抑制剂公开了抑制细胞内转运，细胞表面表达和分泌粘蛋白和O-糖基化糖蛋白的方法，而不影响O-糖基化处理。

9. 发明授权

US06916649B2 UDP-galactose: β-N-acetyl-glucosamine β-1,4-galactosyltransferase, β4Gal-T2 失效
标题翻译： UDP-半乳糖：β-N-乙酰葡糖胺β-1,4-半乳糖基转移酶，β4Gal-T2
公开(公告)号：US06916649B2
公开(公告)日：2005-07-12
申请号：US10132652
申请日：2002-04-24
申请人： Henrik Clausen , Eric Paul Bennett
发明人： Henrik Clausen , Eric Paul Bennett
IPC分类号： C12N9/10 , C12N1/12 , C07H21/04 , C12N1/20 , C12N5/00
CPC分类号： C12N9/1051
摘要： A novel gene defining a novel enzyme in the UDP-D-galactose: b-N-acetyl-glucosamine β-1,4-galactosyltransferase family, termed β4Gal-T2, with unique enzymatic properties is disclosed. The enzymatic activity of β4Gal-T2 is shown to be distinct from that of previously identified enzymes of this gene family. The invention discloses isolated DNA molecules and DNA constructs encoding β4Gal-T2 and derivatives thereof by way of amino acid deletion, substitution or insertion exhibiting β4Gal-T2 activity, as well as cloning and expression vectors including such DNA, cells transfected with the vectors, and recombinant methods for providing β4Gal-T2. The enzyme β4Gal-T2 and β4Gal-T2-active derivatives thereof are disclosed, in particular soluble derivatives comprising the catalytically active domain of β4Gal-T2. Further, the invention discloses methods of obtaining β-1,4-galactosyl glycosylated saccharides, glycopeptides or glycoproteins by use of an enzymically active β4Gal-T2 protein or fusion protein thereof or by using cells stably transfected with a vector including DNA encoding an enzymatically active β4Gal-T2 protein as an expression system for recombinant production of such glycopeptides or glycoproteins. Also a method for the identification of DNA sequence variations in the β4Gal-T2 gene by isolating DNA from a patient, amplifying β4Gal-T2-coding exons by PCR, and detecting the presence of DNA sequence variation, are disclosed.
摘要翻译：公开了一种在UDP-D-半乳糖：b-N-乙酰葡糖胺β-1,4-半乳糖基转移酶家族中定义新的酶的新基因，称为β4Gal-T2，具有独特的酶学性质。 β4Gal-T2的酶活性显示与以前鉴定的该基因家族酶的活性不同。本发明公开了分离的DNA分子和编码β4Gal-T2的DNA构建体及其衍生物，通过氨基酸缺失，取代或插入显示β4Gal-T2活性，以及克隆和表达载体，包括这种DNA，用载体转染的细胞，以及提供β4Gal-T2的重组方法。公开了酶β4Gal-T2和β4Gal-T2-活性衍生物，特别是包含β4Gal-T2的催化活性结构域的可溶性衍生物。此外，本发明公开了通过使用酶活性β4Gal-T2蛋白或其融合蛋白或通过使用包含编码酶活性的DNA的载体稳定转染的细胞获得β-1,4-半乳糖基糖基化糖，糖肽或糖蛋白的方法 β4Gal-T2蛋白作为重组生产这种糖肽或糖蛋白的表达系统。还公开了通过从患者中分离DNA，通过PCR扩增β4Gal-T2编码外显子并检测DNA序列变异的存在来鉴定β4Gal-T2基因中的DNA序列变异的方法。

10. 发明申请

US20110237782A1 METHODS FOR GLYCO-ENGINEERING PLANT CELLS FOR CONTROLLED HUMAN O-GLYCOSYLATION 有权
标题翻译：用于控制人类O-糖蛋白化的植物工程细胞的方法
公开(公告)号：US20110237782A1
公开(公告)日：2011-09-29
申请号：US13070248
申请日：2011-03-23
申请人： Zhang Yang , Damian Paul Drew , Emma Adhiambo Arigi , Peter Ulvskov , Steven B. Levery , Eric Paul Bennett , Henrik Clausen , Brent Larsen Petersen
发明人： Zhang Yang , Damian Paul Drew , Emma Adhiambo Arigi , Peter Ulvskov , Steven B. Levery , Eric Paul Bennett , Henrik Clausen , Brent Larsen Petersen
IPC分类号： C07K14/00 , C12N15/82 , C12P21/00
CPC分类号： C12N15/8257 , C07K14/4727 , C12N15/8245 , C12P21/005
摘要： This invention discloses the development of a novel platform for recombinant production of bioactive glycoproteins and cancer specific vaccines in plants. Plants and plant cell cultures have been humanized with respect to human mucin-type protein O-glycosylation. A panel of plant cell factories for production of recombinant glycoproteins with designed human O-glycosylation, including an improved cancer vaccine candidate, has been developed. The platform provides basis for i) production of an essentially unlimited array of O-glycosylated human glycoprotein therapeutics, such as human interferon α2B and podoplanin, and ii) for further engineering of additional cancer specific O-glycans on glycoproteins of therapeutical value. Currently, mammalian cells are required for human O-glycosylation, but plants offer a unique cell platform for engineering O-glycosylation since they do not perform human type O-glycosylation. Introduction of O-glycosylation into plant cells requires i) that wild-type plant cells do not modify the target peptide substrates and ii) that the appropriate enzymes and substrates are introduced into of plant cells such that O-glycosylation in the secretory pathway proceed and the glycosylated peptide substrates are preferentially exported to the exterior of the cell or accumulated in the cell. In this invention i) the integrity of transiently and stably expressed ‘mucin’ type target peptides in plants cells has been determined and ii) mucin-type O-glycosylation has been established in plants by transient and stable introduction of a Pseudomonas aeruginosa C4-epimerase, the human polypeptide GalNAc-transferases T2 and T4 (GalNAc-T2 and T4) and various human target peptides or proteins. In the present invention GalNAc-T2 and -T4 have been used to produce a Tn cancer glycoform of MUC1.
摘要翻译：本发明公开了在植物中重组生物活性糖蛋白和癌特异性疫苗重组生产的新平台的开发。植物和植物细胞培养物相对于人粘蛋白型蛋白O-糖基化已被人源化。已经开发了一组用于生产具有设计的人O-糖基化的重组糖蛋白的植物细胞工厂，包括改进的癌症疫苗候选物。该平台为i）生产基本上无限量的O-糖基化的人类糖蛋白治疗剂例如人干扰素α2B和podoplanin的基础，以及ii）用于在治疗价值的糖蛋白上进一步工程化另外的癌症特异性O-聚糖。目前，哺乳动物细胞是人O-糖基化所必需的，但由于它们不进行人类O-糖基化，植物提供了独特的用于工程化O-糖基化的细胞平台。将O-糖基化引入到植物细胞中需要i）野生型植物细胞不修饰靶肽底物，和ii）将合适的酶和底物引入植物细胞，使分泌途径中的O-糖基化进行，糖基化肽底物优选地输出到细胞的外部或者积聚在细胞中。在本发明中，i）已经确定了植物细胞中瞬时稳定表达的“粘蛋白”型靶肽的完整性，并且ii）通过暂时稳定地引入铜绿假单胞菌C4差向异构酶，在植物中建立了粘蛋白型O-糖基化，人多肽GalNAc-转移酶T2和T4（GalNAc-T2和T4）和各种人靶肽或蛋白质。在本发明中，已经使用GalNAc-T2和-T4来产生MUC1的Tn癌糖蛋白。

你已经成功收藏专利！

检索式保存成功!

IPRDB

热门服务

关于我们

友情链接

联系方式