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1. 发明授权

US07767415B2 Compositions and methods for modifying blood cell carbohydrates 有权
标题翻译：使用具有独特的底物特异性和动力学性质的α-正乙酰半乳糖胺酶和α-半乳糖苷酶对血型a，b和ab血细胞进行酶转化
公开(公告)号：US07767415B2
公开(公告)日：2010-08-03
申请号：US10251271
申请日：2002-09-20
申请人： Henrik Clausen , Humberto de la Vega , Cheryl Hill , Qiyong Peter Liu
发明人： Henrik Clausen , Humberto de la Vega , Cheryl Hill , Qiyong Peter Liu
IPC分类号： C12P21/06
CPC分类号： C12N5/0641 , C12N9/2402 , C12N9/2465 , C12Q1/34 , C12R1/465 , C12Y302/01022 , C12Y302/01049
摘要： This invention relates to enzymatic removal of type A and B antigens from blood group A, B, and AB reactive cells in blood products, and thereby converting these to non-A and non-B reactive cells. The invention further relates to using unique αN-acetylgalactosaminidases and α-galactosidases with superior kinetic properties for removing the immunodominant monosaccharides of the blood group A and B antigens and improved performance in enzymatic conversion of red blood cells. The preferred unique α-N-acetylgalactosaminidases and α-galactosidases exhibit the following characteristics: (i) exclusive, preferred or no less than 10% substrate specificity for the type A and B branched polysaccharide structures relative to measurable activity with simple mono- and disaccharide structures and aglycon derivatives hereof; (ii) optimal performance at neutral pH with blood group oligosaccharides and in enzymatic conversion of cells; and (iii) a favorable kinetic constant Km with mono- and oligosaccharide substrates. The conversion methods of the invention use significantly lower amounts of recombinant glycosidase enzymes than previous and result in complete sero-conversion of all blood group A and B red cells.
摘要翻译：本发明涉及从血型A，B，AB型反应性细胞中分离A型和B型抗原，从而将其转化为非A型和非B型细胞。本发明还涉及使用独特的αN-乙酰半乳糖胺酶和具有优异动力学性质的α-半乳糖苷酶来除去血型A和B抗原的免疫优势单糖，并提高红细胞酶促转化的表现。优选的独特的α-N-乙酰半乳糖胺酶和α-半乳糖苷酶具有以下特征：（i）相对于简单的单糖和二糖的可测量的活性，对于A型和B型支链多糖结构，独特的，优选的或不低于10％的底物特异性结构和糖苷配基衍生物; （ii）在中性pH下用血型寡糖和细胞酶促转化的最佳性能; 和（iii）具有单糖和寡糖底物的有利的动力学常数Km。本发明的转化方法比以前使用显着更低量的重组糖苷酶，并导致所有血型A和B红细胞的完全血清转化。

2. 发明申请

US20120202273A1 Enzymatic Conversion Of Blood Group A, B, And AB Red Blood Cells Using Alpha-N-Acetylgalactosaminidases and Alpha-Galactosidases With Unique Substrate Specificities And Kinetic Properties 有权
标题翻译：使用具有独特底物特异性和动力学性质的α-N-乙酰半乳糖胺酶和α-半乳糖苷酶对A组，B组和AB组血液细胞进行酶转化
公开(公告)号：US20120202273A1
公开(公告)日：2012-08-09
申请号：US13205588
申请日：2011-08-08
申请人： Henrik Clausen , Humberto de la Vega , Chery Hill , Qiyong Peter Liu
发明人： Henrik Clausen , Humberto de la Vega , Chery Hill , Qiyong Peter Liu
IPC分类号： C12N1/20
CPC分类号： C12N5/0641 , C12N9/2402 , C12N9/2465 , C12Q1/34 , C12R1/465 , C12Y302/01022 , C12Y302/01049
摘要： This invention relates to enzymatic removal of type A and B antigens from blood group A, B, and AB reactive cells in blood products, and thereby converting these to non-A and non-B reactive cells. The invention further relates to using unique α-N-acetylgalactosaminidases and α-galactosidases with superior kinetic properties for removing the immunodominant monosaccharides of the blood group A and B antigens and improved performance in enzymatic conversion of red blood cells. The preferred unique α-N-acetylgalactosaminidases and α-galactosidases exhibit the following characteristics: (i) exclusive, preferred or no less than 10% substrate specificity for the type A and B branched polysaccharide structures relative to measurable activity with simple mono- and disaccharide structures and aglycon derivatives hereof; (ii) optimal performance at neutral pH with blood group oligosaccharides and in enzymatic conversion of cells; and (iii) a favorable kinetic constant Km with mono- and oligosaccharide substrates. The conversion methods of the invention use significantly lower amounts of recombinant glycosidase enzymes than previous and result in complete sero-conversion of all blood group A and B red cells.
摘要翻译：本发明涉及从血型A，B，AB型反应性细胞中分离A型和B型抗原，从而将其转化为非A型和非B型细胞。本发明还涉及使用独特的α-N-乙酰半乳糖胺酶和具有优异动力学性质的α-半乳糖苷酶，用于去除血型A和B抗原的免疫优势单糖，并提高了红细胞酶促转化的表现。优选的独特的α-N-乙酰半乳糖胺酶和α-半乳糖苷酶具有以下特征：（i）相对于简单的单糖和二糖的可测量的活性，对A型和B型支链多糖结构的排列，优选或不低于10％的底物特异性结构和糖苷配基衍生物; （ii）在中性pH下用血型寡糖和细胞酶促转化的最佳性能; 和（iii）具有单糖和寡糖底物的有利的动力学常数Km。本发明的转化方法比以前使用显着更低量的重组糖苷酶，并导致所有血型A和B红细胞的完全血清转化。

3. 发明申请

US20110014170A1 NOVEL ALPHA-GALACTOSIDASES 有权
标题翻译：新的阿尔法胶质糖
公开(公告)号：US20110014170A1
公开(公告)日：2011-01-20
申请号：US12709070
申请日：2010-02-19
申请人： Qiyong Peter LIU , Henrik Clausen
发明人： Qiyong Peter LIU , Henrik Clausen
IPC分类号： A61K35/18 , A61P7/00 , C12N5/078 , C12N9/24
CPC分类号： C12N5/0654 , A61K35/32 , A61K35/35 , A61L27/3687 , A61L27/3695 , C12N5/0652 , C12N9/2465 , C12N2501/73 , C12N2506/09 , C12N2506/13 , C12N2506/1315 , C12N2506/14 , C12N2506/25 , C12Y302/01022
摘要： This invention relates to novel α-galactosidases for the enzymatic removal of the immunodominant monosaccharides on blood products and tissues. Specifically this invention provides a novel family of α3 glycosidases, used for the enzymatic removal of type B antigens from blood group B and AB reactive blood products, and the Galili antigen from non-human animal tissues, thereby converting these to non-immunogenic cells and tissues suitable for transplantation.
摘要翻译：本发明涉及用于在血液制品和组织上酶促去除免疫优势单糖的新型α-半乳糖苷酶。具体地说，本发明提供了一种新型的α3糖苷酶家族，用于从血型B和AB反应性血液制品中酶切除B型抗原，从非人动物组织中提取Galectin抗原，从而将其转化为非免疫原性细胞，适合移植的组织。

4. 发明授权

US08883478B2 Methods of preparing tissues for xenotransplantation using α-galactosidases 有权
标题翻译：使用α-半乳糖苷酶制备异种移植组织的方法
公开(公告)号：US08883478B2
公开(公告)日：2014-11-11
申请号：US12709070
申请日：2010-02-19
申请人： Qiyong Peter Liu , Henrik Clausen
发明人： Qiyong Peter Liu , Henrik Clausen
IPC分类号： C12Q1/34 , C12N9/40 , C12P21/00 , C07K14/00
CPC分类号： C12N5/0654 , A61K35/32 , A61K35/35 , A61L27/3687 , A61L27/3695 , C12N5/0652 , C12N9/2465 , C12N2501/73 , C12N2506/09 , C12N2506/13 , C12N2506/1315 , C12N2506/14 , C12N2506/25 , C12Y302/01022
摘要： This invention relates to novel α-galactosidases for the enzymatic removal of the immunodominant monosaccharides on blood products and tissues. Specifically this invention provides a novel family of α3 glycosidases, used for the enzymatic removal of type B antigens from blood group B and AB reactive blood products, and the Galili antigen from non-human animal tissues, thereby converting these to non-immunogenic cells and tissues suitable for transplantation.
摘要翻译：本发明涉及用于在血液制品和组织上酶促去除免疫优势单糖的新型α-半乳糖苷酶。具体地说，本发明提供了一种新型的α3糖苷酶家族，用于从血型B和AB反应性血液制品中酶切除B型抗原，从非人动物组织中提取Galectin抗原，从而将其转化为非免疫原性细胞，适合移植的组织。

5. 发明申请

US20110045569A1 ENZYMATIC CONVERSION OF BLOOD GROUP A, B, AND AB RED BLOOD CELLS USING ALPHA-N- ACETYLGALACTOSAMINIDASES AND ALPHA-GALACTOSIDASES WITH UNIQUE SUBSTRATE SPECIFICITIES AND KINETIC PROPERTIES 失效
标题翻译：使用ALPHA-N-乙酰胆碱酯酶和ALPHA-GALACTOSIDASES具有独特的底物特异性和动力学特性的组A，B和AB红血细胞的酶转化
公开(公告)号：US20110045569A1
公开(公告)日：2011-02-24
申请号：US12789946
申请日：2010-05-28
申请人： Henrik Clausen , Humberto De La Vega , Cheryl Hill , Qiyong Peter Liu
发明人： Henrik Clausen , Humberto De La Vega , Cheryl Hill , Qiyong Peter Liu
IPC分类号： C12N9/40 , C12N1/20
CPC分类号： C12N5/0641 , C12N9/2402 , C12N9/2465 , C12Q1/34 , C12R1/465 , C12Y302/01022 , C12Y302/01049
摘要： This invention relates to enzymatic removal of type A and B antigens from blood group A, B, and AB reactive cells in blood products, and thereby converting these to non-A and non-B reactive cells. The invention further relates to using unique α-N-acetylgalactosaminidases and α-galactosidases with superior kinetic properties for removing the immunodominant monosaccharides of the blood group A and B antigens and improved performance in enzymatic conversion of red blood cells. The preferred unique α-N-acetylgalactosaminidases and α-galactosidases exhibit the following characteristics: (i) exclusive, preferred or no less than 10% substrate specificity for the type A and B branched polysaccharide structures relative to measurable activity with simple mono- and disaccharide structures and aglycon derivatives hereof; (ii) optimal performance at neutral pH with blood group oligosaccharides and in enzymatic conversion of cells; and (iii) a favorable kinetic constant Km with mono- and oligosaccharide substrates. The conversion methods of the invention use significantly lower amounts of recombinant glycosidase enzymes than previous and result in complete sero-conversion of all blood group A and B red cells.
摘要翻译：本发明涉及从血型A，B，AB型反应性细胞中分离A型和B型抗原，从而将其转化为非A型和非B型细胞。本发明还涉及使用独特的α-N-乙酰半乳糖胺酶和具有优异动力学性质的α-半乳糖苷酶，用于去除血型A和B抗原的免疫优势单糖，并提高了红细胞酶促转化的表现。优选的独特的α-N-乙酰半乳糖胺酶和α-半乳糖苷酶具有以下特征：（i）相对于简单的单糖和二糖的可测量的活性，对于A型和B型支链多糖结构，独特的，优选的或不低于10％的底物特异性结构和糖苷配基衍生物; （ii）在中性pH下用血型寡糖和细胞酶促转化的最佳性能; 和（iii）具有单糖和寡糖底物的有利的动力学常数Km。本发明的转化方法比以前使用显着更低量的重组糖苷酶，并导致所有血型A和B红细胞的完全血清转化。

6. 发明授权

US08697411B2 Streptomyces griseoplanus comprising an α-galactosidase for removing immunodominant α-galactose monosaccharides from blood group B or AB reactive cells 有权
标题翻译：包含α-半乳糖苷酶的灰色链霉菌，用于从血型B或AB反应性细胞中除去免疫优势的α-半乳糖单糖
公开(公告)号：US08697411B2
公开(公告)日：2014-04-15
申请号：US13205588
申请日：2011-08-08
申请人： Henrik Clausen , Humberto de la Vega , Cheryl Hill , Qiyong Peter Liu
发明人： Henrik Clausen , Humberto de la Vega , Cheryl Hill , Qiyong Peter Liu
IPC分类号： C12N9/24
CPC分类号： C12N5/0641 , C12N9/2402 , C12N9/2465 , C12Q1/34 , C12R1/465 , C12Y302/01022 , C12Y302/01049
摘要： This invention relates to enzymatic removal of type A and B antigens from blood group A, B, and AB reactive cells in blood products, and thereby converting these to non-A and non-B reactive cells. The invention further relates to using unique α-N-acetylgalactosaminidases and α-galactosidases with superior kinetic properties for removing the immunodominant monosaccharides of the blood group A and B antigens and improved performance in enzymatic conversion of red blood cells. The preferred unique α-N-acetylgalactosaminidases and α-galactosidases exhibit the following characteristics: (i) exclusive, preferred or no less than 10% substrate specificity for the type A and B branched polysaccharide structures relative to measurable activity with simple mono- and disaccharide structures and aglycon derivatives hereof; (ii) optimal performance at neutral pH with blood group oligosaccharides and in enzymatic conversion of cells; and (iii) a favorable kinetic constant Km with mono- and oligosaccharide substrates. The conversion methods of the invention use significantly lower amounts of recombinant glycosidase enzymes than previous and result in complete sero-conversion of all blood group A and B red cells.
摘要翻译：本发明涉及从血型A，B，AB型反应性细胞中分离A型和B型抗原，从而将其转化为非A型和非B型细胞。本发明还涉及使用独特的α-N-乙酰半乳糖胺酶和具有优异动力学性质的α-半乳糖苷酶，用于去除血型A和B抗原的免疫优势单糖，并提高了红细胞酶促转化的表现。优选的独特的α-N-乙酰半乳糖胺酶和α-半乳糖苷酶具有以下特征：（i）相对于简单的单糖和二糖的可测量的活性，对于A型和B型支链多糖结构，独特的，优选的或不低于10％的底物特异性结构和糖苷配基衍生物; （ii）在中性pH下用血型寡糖和细胞酶促转化的最佳性能; 和（iii）具有单糖和寡糖底物的有利的动力学常数Km。本发明的转化方法比以前使用显着更低量的重组糖苷酶，并导致所有血型A和B红细胞的完全血清转化。

7. 发明授权

US07993896B2 Streptomyces griseoplanus α-galactosidases for removing immunodominant α-galactose monosaccharides from blood group B or AB reactive cells 失效
标题翻译：用于从血型B或AB反应性细胞中去除免疫显性α-半乳糖单糖的灰色链霉菌α-半乳糖苷酶
公开(公告)号：US07993896B2
公开(公告)日：2011-08-09
申请号：US12789946
申请日：2010-05-28
申请人： Henrik Clausen , Humberto de la Vega , Cheryl Hill , Qiyong Peter Liu
发明人： Henrik Clausen , Humberto de la Vega , Cheryl Hill , Qiyong Peter Liu
IPC分类号： C12N9/24
CPC分类号： C12N5/0641 , C12N9/2402 , C12N9/2465 , C12Q1/34 , C12R1/465 , C12Y302/01022 , C12Y302/01049
摘要： This invention relates to enzymatic removal of type A and B antigens from blood group A, B, and AB reactive cells in blood products, and thereby converting these to non-A and non-B reactive cells. The invention further relates to using unique α-N-acetylgalactosaminidases and α-galactosidases with superior kinetic properties for removing the immunodominant monosaccharides of the blood group A and B antigens and improved performance in enzymatic conversion of red blood cells. The preferred unique α-N-acetylgalactosaminidases and α-galactosidases exhibit the following characteristics: (i) exclusive, preferred or no less than 10% substrate specificity for the type A and B branched polysaccharide structures relative to measurable activity with simple mono- and disaccharide structures and aglycon derivatives hereof; (ii) optimal performance at neutral pH with blood group oligosaccharides and in enzymatic conversion of cells; and (iii) a favorable kinetic constant Km with mono- and oligosaccharide substrates. The conversion methods of the invention use significantly lower amounts of recombinant glycosidase enzymes than previous and result in complete sero-conversion of all blood group A and B red cells.
摘要翻译：本发明涉及从血型A，B，AB型反应性细胞中分离A型和B型抗原，从而将其转化为非A型和非B型细胞。本发明还涉及使用独特的α-N-乙酰半乳糖胺酶和具有优异动力学性质的α-半乳糖苷酶，用于去除血型A和B抗原的免疫优势单糖，并提高了红细胞酶促转化的表现。优选的独特的α-N-乙酰半乳糖胺酶和α-半乳糖苷酶具有以下特征：（i）相对于简单的单糖和二糖的可测量的活性，对于A型和B型支链多糖结构，独特的，优选的或不低于10％的底物特异性结构和糖苷配基衍生物; （ii）在中性pH下用血型寡糖和细胞酶促转化的最佳性能; 和（iii）具有单糖和寡糖底物的有利的动力学常数Km。本发明的转化方法比以前使用显着更低量的重组糖苷酶，并导致所有血型A和B红细胞的完全血清转化。

8. 发明授权

US07951552B2 Method of preparing non-human tissues for xenotransplantation using α-galactosidase 有权
标题翻译：使用α-半乳糖苷酶制备用于异种移植的非人组织的方法
公开(公告)号：US07951552B2
公开(公告)日：2011-05-31
申请号：US12091818
申请日：2006-10-31
申请人： Qiyong Peter Liu , Henrik Clausen
发明人： Qiyong Peter Liu , Henrik Clausen
IPC分类号： A61K35/36 , C12Q1/34 , C12N9/40 , C12P21/00 , C07K14/00
CPC分类号： C12N5/0654 , A61K35/32 , A61K35/35 , A61L27/3687 , A61L27/3695 , C12N5/0652 , C12N9/2465 , C12N2501/73 , C12N2506/09 , C12N2506/13 , C12N2506/1315 , C12N2506/14 , C12N2506/25 , C12Y302/01022
摘要： This invention relates to novel α-galactosidases for the enzymatic removal of the immunodominant monosaccharides on blood products and tissues. Specifically this invention provides a novel family of α3 glycosidases, used for the enzymatic removal of type B antigens from blood group B and AB reactive blood products, and the GaIiIi antigen from non-human animal tissues, thereby converting these to non-immunogenic cells and tissues suitable for transplantation.
摘要翻译：本发明涉及用于在血液制品和组织上酶促去除免疫优势单糖的新型α-半乳糖苷酶。具体地说，本发明提供了一种新型的α3糖苷酶家族，其用于从血型B和AB反应性血液制品中酶切除B型抗原，以及来自非人动物组织的GaIiI抗原，从而将其转化为非免疫原性细胞，适合移植的组织。

9. 发明授权

US09029331B2 Glycopegylated granulocyte colony stimulating factor 有权
标题翻译：糖基化的粒细胞集落刺激因子
公开(公告)号：US09029331B2
公开(公告)日：2015-05-12
申请号：US11794560
申请日：2006-01-10
申请人： Shawn DeFrees , Henrik Clausen , David A. Zopf , Caryn Bowe , Susann Taudte , Michael Felo , Walter S. Willett
发明人： Shawn DeFrees , Henrik Clausen , David A. Zopf , Caryn Bowe , Susann Taudte , Michael Felo , Walter S. Willett
IPC分类号： A61K38/14 , A61K38/16 , A61K38/22 , C07K14/575 , C07K14/535 , C07K1/18 , C07K1/107 , A61K47/48 , A61K38/00
CPC分类号： C07K14/535 , A61K38/00 , A61K47/549 , A61K47/60 , A61K47/62 , C07K1/1077 , C07K1/18
摘要： The invention relates to methods of preparing and purifying conjugates between Granulocyte Colony Stimulating Factor and PEG moieties. The conjugates are linked via an intact glycosyl linking group that is interposed between and covalently attached to the peptide and the modifying group. The conjugates are purified using various chromatography methods.
摘要翻译：本发明涉及制备和纯化粒细胞集落刺激因子和PEG部分之间的缀合物的方法。缀合物经由完整的糖基连接基团连接，该基团介于并共价连接到肽和修饰基团之间。使用各种色谱法纯化缀合物。

10. 发明授权

US09024110B2 Methods for glyco-engineering plant cells for controlled human O-glycosylation 有权
标题翻译：糖酵解植物细胞用于受控人O-糖基化的方法
公开(公告)号：US09024110B2
公开(公告)日：2015-05-05
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 Bennett , Henrik Clausen , Brent Larsen Petersen
发明人： Zhang Yang , Damian Paul Drew , Emma Adhiambo Arigi , Peter Ulvskov , Steven B. Levery , Eric Bennett , Henrik Clausen , Brent Larsen Petersen
IPC分类号： C12N15/82 , C12N15/12 , C12N15/30 , C12N15/31 , C12N9/10 , C12N9/90 , C12P19/00 , C12P19/44 , A61K38/47 , C12N1/13 , C07K14/47 , 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。

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