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1. 发明公开

EP3017057A1 PROCESS FOR THE MONO- AND BI-SIALYLATION OF GLYCOPROTEINS EMPLOYING N-TERMINALLY TRUNCATED BETA-GALACTOSIDE ALPHA-2,6-SIALYLTRANSFERASE MUTANTS 有权
标题翻译： MONO的方法和糖蛋白WITH BISIALYLIERUNG N-末端CONDENSEDβ-半乳糖苷-ALPHA-2,6-唾液酸转移酶突变体
公开(公告)号：EP3017057A1
公开(公告)日：2016-05-11
申请号：EP14736379.0
申请日：2014-07-03
申请人： F.Hoffmann-La Roche AG , Roche Diagnostics GmbH
发明人： CZABANY, Tibor , ENGEL, Alfred , GREIF, Michael , JUNG, Christine , LULEY, Christiane , MALIK, Sebastian , MUELLER, Rainer , NIDETZKY, Bernd , RIBITSCH, Doris , SCHMOELZER, Katharina , SCHWAB, Helmut , SOBEK, Harald , SUPPMANN, Bernhard , THOMANN, Marco , ZITZENBACHER, Sabine
IPC分类号： C12P19/18 , C12P21/00 , C12N9/10 , C07K16/00
摘要： The present disclosure is directed to the use of certain glycosyltransferase variants having N-terminal truncation deletions. It was found that the combination of two different truncation variants of human β-galactoside-α-2,6-sialyltransferase I (hST6Gal-I) exhibited different specific sialyltransferase enzymatic activities. In one example, under conditions wherein the first variant Δ89 hST6Gal-I catalyzed formation of bi-sialylated target molecules the second variant Δ108 hST6Gal-I catalyzed formation of mono-sialylated target molecules. Thus, disclosed are variants of mammalian glycosyltransferase, nucleic acids encoding the same, methods and means for recombinantly producing the variants of mammalian glycosyltransferase and use thereof, particularly for sialylating in a quantitatively controlled manner terminal acceptor groups of glycan moieties being part of glycoproteins such as immunoglobulins.

2. 发明公开

EP3017041A1 N-TERMINALLY TRUNCATED GLYCOSYLTRANSFERASES 审中-公开
标题翻译： N-末端TRUNKIERTE GLYKOSYLTRANSFERASEN
公开(公告)号：EP3017041A1
公开(公告)日：2016-05-11
申请号：EP14735569.7
申请日：2014-07-03
申请人： F.Hoffmann-La Roche AG , Roche Diagnostics GmbH
发明人： CZABANY, Tibor , ENGEL, Alfred , GREIF, Michael , JUNG, Christine , LULEY, Christiane , MALIK, Sebastian , MUELLER, Rainer , NIDETZKY, Bernd , RIBITSCH, Doris , SCHMOELZER, Katharina , SCHWAB, Helmut , SOBEK, Harald , SUPPMANN, Bernhard , THOMANN, Marco , ZITZENBACHER, Sabine
IPC分类号： C12N9/10
摘要： The present disclosure is directed to glycosyltransferase variants having N-terminal truncation deletions. Contrary to previous findings certain truncations comprising the conserved amino acid motif ("QVWxKDS") were found to be compatible with glycosyltransferase enzymatic activity, particularly in a human sialyltransferase (hST6Gal-I). Thus, disclosed are variants of mammalian glycosyltransferase, nucleic acids encoding the same, methods and means for recombinantly producing the variants of mammalian glycosyltransferase and use thereof, particularly for sialylating terminal acceptor groups of glycan moieties being part of glycoproteins such as immunoglobulins.
摘要翻译：本公开涉及具有N-末端截短缺失的糖基转移酶变体。与以前的发现相反，发现包含保守氨基酸基序（“QVWxKDS”）的某些截短与糖基转移酶酶活性相容，特别是在人唾液酸转移酶（hST6Gal-1）中。因此，所公开的是哺乳动物糖基转移酶的变体，编码它们的核酸，用于重组产生哺乳动物糖基转移酶变体的方法和手段及其用途，特别是作为糖蛋白的部分的聚糖末端受体基团（如免疫球蛋白）的唾液酸化。

3. 发明公开

EP3004368A1 QUANTITATIVE CONTROL OF SIALYLATION 有权
标题翻译：定量管制SIALYIERUNG
公开(公告)号：EP3004368A1
公开(公告)日：2016-04-13
申请号：EP14725130.0
申请日：2014-05-16
申请人： F.Hoffmann-La Roche AG , Roche Diagnostics GmbH
发明人： ENGEL, Alfred , GREIF, Michael , JUNG, Christine , MALIK, Sebastian , MUELLER, Rainer , SOBEK, Harald , SUPPMANN, Bernhard , THOMANN, Marco
IPC分类号： C12P21/00 , C12P19/18 , A61K39/395
摘要： The present disclosure is directed to the use of certain glycosyltransferase variants having N-terminal truncation deletions. Contrary to previous findings certain truncations were found to exhibit sialidase enzymatic activity, particularly a variant of human sialyltransferase (hST6Gal-I) with a truncation deletion involving the first 89 N-terminal amino acids of the respective wild-type polypeptide. A fundamental finding documented in the present disclosure is that there exists a variant of this enzyme which is capable of catalyzing transfer of a glycosyl moiety as well as hydrolysis thereof. Thus, disclosed is a specific exemplary variant of mammalian glycosyltransferase, nucleic acids encoding the same, methods and means for recombinantly producing the variant of mammalian glycosyltransferase and use thereof, particularly for sialylating in a quantitatively controlled manner terminal acceptor groups of glycan moieties being part of glycoproteins such as immunoglobulins.

5. 发明授权

EP1144653B1 THERMOSTABLE ENZYME PROMOTING THE FIDELITY OF THERMOSTABLE DNA POLYMERASES- FOR IMPROVEMENT OF NUCLEIC ACID SYNTHESIS AND AMPLIFICATION IN VITRO 有权
标题翻译：热稳定性酶WHICH准确性THERMO STABLE DNA聚合酶INCREASES - 以改善体外和扩增核酸的合成
公开(公告)号：EP1144653B1
公开(公告)日：2006-08-09
申请号：EP00969312.8
申请日：2000-09-27
申请人： Roche Diagnostics GmbH
发明人： ANKENBAUER, Waltraud , LAUE, Frank , SOBEK, Harald , GREIF, Michael
IPC分类号： C12N15/55 , C12N15/54 , C12N15/10 , C12N9/22 , C12N9/12 , C12Q1/68 , C12P19/34
CPC分类号： C12N15/10 , C12N9/1252 , C12N9/22 , C12Q1/686 , C12Q2521/101
摘要： A purified thermostable enzyme is derived from the thermophilic archaebacterium Archaeoglobus fulgidus. The enzyme can be native or recombinant, is stable under PCR conditions and exhibits double strand specific exonuclease activity. It is a 3'-5' exonuclease and cleaves to produce 5'-mononucleotides. Thermostable exonucleases are useful in many recombinant DNA techniques, in combination with a thermostable DNA polymerase like Tag especially for nucleic acid amplification by the polymerase chain reaction (PCR).

6. 发明公开

EP3237608A1 CMP-DEPENDENT SIALIDASE ACTIVITY 有权
标题翻译： CMP依赖性唾液酸酶活性
公开(公告)号：EP3237608A1
公开(公告)日：2017-11-01
申请号：EP15817301.3
申请日：2015-12-21
申请人： F. Hoffmann-La Roche AG , Roche Diagnostics GmbH
发明人： SOBEK, Harald , GREIF, Michael , THOMANN, Marco , MALIK, Sebastian
IPC分类号： C12N9/10 , C12P19/44
摘要： The present disclosure is directed to the properties of certain glycosyltransferase variants having N-terminal truncation deletions or internal deletions. Any of the mutants disclosed in here exhibit α-2,6-sialyltransferase enzymatic activity in the presence of CMP-activated sialic acid as co-substrate, and in the presence of a suitable acceptor site. A fundamental finding documented in the present disclosure is that suchs enzyme are not only capable of catalyzing transfer of a sialidyl moiety but they are also capable of catalyzing hydrolytic cleavage of terminally bound sialic acid from a glycan.
摘要翻译：本公开涉及具有N端截短缺失或内部缺失的某些糖基转移酶变体的性质。此处公开的任何突变体在CMP-活化的唾液酸作为共底物并存在合适的受体位点的情况下表现出α-2,6-唾液酸转移酶酶活性。本公开中记载的基本发现是，这样的酶不仅能够催化唾液酸基部分的转移，而且它们还能够催化从聚糖水解切割末端结合的唾液酸。特别是发现在胞苷5'-单磷酸（CMP）存在下糖基转移酶活性受到抑制，并且唾液酸酶活性受到刺激。发现唾液酸酶活性依赖于参考（野生型）hST6Gal-1多肽的多肽序列中特定氨基酸片段（位置90至108）的存在。发现N-末端截短变体中该序列部分的缺失可消除唾液酸酶活性，特别是在存在和不存在CMP时。因此，公开了使用本文记载的CMP介导的反馈调节的组合物，用途和方法。

7. 发明授权

EP3237608B1 CMP-DEPENDENT SIALIDASE ACTIVITY 有权
公开(公告)号：EP3237608B1
公开(公告)日：2019-10-09
申请号：EP15817301.3
申请日：2015-12-21
申请人： F. Hoffmann-La Roche AG , Roche Diagnostics GmbH
发明人： SOBEK, Harald , GREIF, Michael , THOMANN, Marco , MALIK, Sebastian
IPC分类号： C12N9/10 , C12P19/44 , C12P21/00

9. 发明公开

EP1144653A3 THERMOSTABLE ENZYME PROMOTING THE FIDELITY OF THERMOSTABLE DNA POLYMERASES- FOR IMPROVEMENT OF NUCLEIC ACID SYNTHESIS AND AMPLIFICATION i IN VITRO /i 审中-公开
标题翻译：可热稳定的酶促进热稳定的DNA聚合物的保质期 - 用于改善体内核酸合成和功能的扩增
公开(公告)号：EP1144653A3
公开(公告)日：2002-02-13
申请号：EP00969312.8
申请日：2000-09-27
申请人： Roche Diagnostics GmbH
发明人： ANKENBAUER, Waltraud , LAUE, Frank , SOBEK, Harald , GREIF, Michael
IPC分类号： C12N15/55 , C12N15/54 , C12N15/10 , C12N9/22 , C12N9/12 , C12Q1/68 , C12P19/34

10. 发明公开

EP1144653A2 THERMOSTABLE ENZYME PROMOTING THE FIDELITY OF THERMOSTABLE DNA POLYMERASES- FOR IMPROVEMENT OF NUCLEIC ACID SYNTHESIS AND AMPLIFICATION i IN VITRO /i 有权
标题翻译：热稳定性酶WHICH准确性THERMO STABLE DNA聚合酶INCREASES - 以改善体外和扩增核酸的合成
公开(公告)号：EP1144653A2
公开(公告)日：2001-10-17
申请号：EP00969312.8
申请日：2000-09-27
申请人： Roche Diagnostics GmbH
发明人： ANKENBAUER, Waltraud , LAUE, Frank , SOBEK, Harald , GREIF, Michael
IPC分类号： C12N15/55 , C12N15/54 , C12N15/10 , C12N9/22 , C12N9/12 , C12Q1/68 , C12P19/34
CPC分类号： C12N15/10 , C12N9/1252 , C12N9/22 , C12Q1/686 , C12Q2521/101
摘要： A purified thermostable enzyme is derived from the thermophilic archaebacterium Archaeoglobus fulgidus. The enzyme can be native or recombinant, is stable under PCR conditions and exhibits double strand specific exonuclease activity. It is a 3'-5' exonuclease and cleaves to produce 5'-mononucleotides. Thermostable exonucleases are useful in many recombinant DNA techniques, in combination with a thermostable DNA polymerase like Tag especially for nucleic acid amplification by the polymerase chain reaction (PCR).

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