公开(公告)号：US12234488B2

公开(公告)日：2025-02-25

申请号：US18068739

申请日：2022-12-20

Applicant: Roche Diagnostics Operations, Inc.

Inventor： Christian Helmut Bell ,  Harald Sobek ,  Heiko Walch ,  Kiyoshi Yasukawa ,  Misato Baba

IPC: C12P19/34 ,  C12N9/12 ,  C12Q1/68

Abstract: The present invention relates to a mutant reverse transcriptase (RT) with increased thermal stability relative to the wildtype, a nucleic acid encoding the mutant RT, a cell comprising the mutant RT or the nucleic acid, a kit comprising the mutant RT, the use of the mutant RT for cDNA synthesis, method for reverse transcription of RNA comprising synthesizing cDNA with the use of the mutant RT and a method for detecting an RNA marker in a sample with the use of the mutant RT.

公开(公告)号：US11618892B2

公开(公告)日：2023-04-04

申请号：US16599407

申请日：2019-10-11

Applicant: Roche Diagnostics Operations, Inc.

Inventor： Christian Helmut Bell ,  Harald Sobek ,  Heiko Walch ,  Kiyoshi Yasukawa ,  Misato Baba

IPC: A61K38/00 ,  C12N9/12 ,  C12Q1/68 ,  C12P19/34

Abstract: The present invention relates to a mutant reverse transcriptase (RT) with increased thermal stability relative to the wildtype, a nucleic acid encoding the mutant RT, a cell comprising the mutant RT or the nucleic acid, a kit comprising the mutant RT, the use of the mutant RT for cDNA synthesis, method for reverse transcription of RNA comprising synthesizing cDNA with the use of the mutant RT and a method for detecting an RNA marker in a sample with the use of the mutant RT.

公开(公告)号：US20160102298A1

公开(公告)日：2016-04-14

申请号：US14970832

申请日：2015-12-16

Applicant: Roche Diagnostics Operations, Inc.

Inventor： Tibor Czabany ,  Alfred Engel ,  Michael Greif ,  Christine Jung ,  Christiane Luley ,  Sebastian Malik ,  Rainer Mueller ,  Bernd Nidetzky ,  Doris Ribitsch ,  Katharina Schmoelzer ,  Helmut Schwab ,  Harald Sobek ,  Bernhard Suppmann ,  Marco Thomann ,  Sabine Zitzenbacher

IPC: C12N9/10

CPC classification number: C12N9/1081 ,  C12N9/1048 ,  C12Y204/99001

Abstract: 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.

Abstract translation: 本公开涉及具有N-末端截短缺失的糖基转移酶变体。 与之前的发现相反，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，公开了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，特别是用作糖蛋白的部分的聚糖末端受体基团，例如免疫球蛋白。

公开(公告)号：US20160032260A1

公开(公告)日：2016-02-04

申请号：US14885727

申请日：2015-10-16

Applicant: ROCHE DIAGNOSTICS OPERATIONS, INC.

Inventor： Harald Sobek ,  Johann-Peter Thalhofer ,  Rainer Mueller ,  Manfred Schmidt ,  Michael Greif ,  Armin Ruf ,  Christian Rudolph

IPC: C12N9/12

CPC classification number: C12N9/1247 ,  C12P19/34 ,  C12Y207/07006

Abstract: The present invention provides improved variants of T7 RNA polymerase by introducing novel mutations which lead to improved thermostability of the enzyme. According to the invention, amino acid substitutions at the positions Val426, Ser633, Val650, Thr654, Ala702, Val795, and combinations thereof are advantageous.

公开(公告)号：US11788108B2

公开(公告)日：2023-10-17

申请号：US17355962

申请日：2021-06-23

Applicant: Roche Diagnostics Operations, Inc.

Inventor： Harald Sobek ,  Michael Greif ,  Marco Thomann ,  Sebastian Malik

IPC: C12P21/00 ,  C12N9/10 ,  C12P19/44 ,  C07K16/00

CPC classification number: C12P21/005 ,  C07K16/00 ,  C12N9/1081 ,  C12P19/44 ,  C12Y204/99001 ,  C12Y301/03 ,  C07K2317/41

Abstract: The properties of certain glycosyltransferase variants having N-terminal truncation deletions or internal deletions are disclosed. Particularly, mutants that 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, are disclosed. A fundamental finding documented in the present disclosure is that enzymes 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.

公开(公告)号：US20200040316A1

公开(公告)日：2020-02-06

申请号：US16599407

申请日：2019-10-11

Applicant: Roche Diagnostics Operations, Inc.

Inventor： Christian Helmut Bell ,  Harald Sobek ,  Heiko Walch ,  Kiyoshi Yasukawa ,  Misato Baba

IPC: C12N9/12 ,  C12Q1/68

Abstract: The present invention relates to a mutant reverse transcriptase (RT) with increased thermal stability relative to the wildtype, a nucleic acid encoding the mutant RT, a cell comprising the mutant RT or the nucleic acid, a kit comprising the mutant RT, the use of the mutant RT for cDNA synthesis, method for reverse transcription of RNA comprising synthesizing cDNA with the use of the mutant RT and a method for detecting an RNA marker in a sample with the use of the mutant RT.

公开(公告)号：US09631233B2

公开(公告)日：2017-04-25

申请号：US14855915

申请日：2015-09-16

Applicant: ROCHE DIAGNOSTICS OPERATIONS, INC.

Inventor： Thomas Metzler ,  Tobias Haslinger ,  Annette Peceny ,  Harald Sobek

IPC: C12P19/34 ,  C12Q1/68 ,  C12N9/12

CPC classification number: C12Q1/686 ,  C12N9/1252 ,  C12Q1/6876

Abstract: The present invention provides methods to obtain dry compositions of reaction compounds that maintain the biological activity of the compounds upon re-solubilization after a certain storage time. Preferably, the dry composition comprises a polymerase, and the dry composition is usable for polymerase chain reaction (PCR) amplification after re-solubilization.

公开(公告)号：US20140134633A1

公开(公告)日：2014-05-15

申请号：US14160453

申请日：2014-01-21

Applicant: ROCHE DIAGNOSTICS OPERATIONS, INC.

Inventor： Ulrike Fischer ,  Michael Greif ,  Harald Sobek ,  Johann-Peter Thalhofer

IPC: C12Q1/68

CPC classification number: C12Q1/686 ,  C12N9/1252 ,  C12Q2527/137 ,  C12Q2521/101

Abstract: The present invention relates to a formulation of a thermostable DNA polymerase which is completely free of detergents and its particular use in real time polymerase chain reaction (PCR). Such a formulation may be obtained if the selected purification method does not require the addition of a detergent at any purification step.

Abstract translation: 本发明涉及完全不含洗涤剂的热稳定性DNA聚合酶的制剂及其在实时聚合酶链式反应（PCR）中的特别用途。 如果所选择的纯化方法不需要在任何纯化步骤中添加洗涤剂，则可以获得这样的制剂。

公开(公告)号：US20230203460A1

公开(公告)日：2023-06-29

申请号：US18068739

申请日：2022-12-20

Applicant: Roche Diagnostics Operations, Inc.

Inventor： Christian Helmut Bell ,  Harald Sobek ,  Heiko Walch ,  Kiyoshi Yasukawa ,  Misato Baba

IPC: C12N9/12 ,  C12Q1/68 ,  C12P19/34

CPC classification number: C12N9/1276 ,  C12Q1/68 ,  C12Y207/07049 ,  C12P19/34

Abstract: The present invention relates to a mutant reverse transcriptase (RT) with increased thermal stability relative to the wildtype, a nucleic acid encoding the mutant RT, a cell comprising the mutant RT or the nucleic acid, a kit comprising the mutant RT, the use of the mutant RT for cDNA synthesis, method for reverse transcription of RNA comprising synthesizing cDNA with the use of the mutant RT and a method for detecting an RNA marker in a sample with the use of the mutant RT.

公开(公告)号：US09481902B2

公开(公告)日：2016-11-01

申请号：US14970734

申请日：2015-12-16

Applicant: Roche Diagnostics Operations, Inc.

Inventor： Tibor Czabany ,  Alfred Engel ,  Michael Greif ,  Christine Jung ,  Christiane Luley ,  Sebastian Malik ,  Rainer Mueller ,  Bernd Nidetzky ,  Doris Ribitsch ,  Katharina Schmoelzer ,  Helmut Schwab ,  Harald Sobek ,  Bernhard Suppmann ,  Marco Thomann ,  Sabine Zitzenbacher

IPC: C07K16/00 ,  C12Q1/00 ,  C12P21/00 ,  C12N9/10

CPC classification number: C12P21/005 ,  C07K16/00 ,  C07K2317/14 ,  C07K2317/24 ,  C07K2317/41 ,  C12N9/1081 ,  C12P19/18 ,  C12Y204/99001

Abstract: 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.

Abstract translation: 本公开涉及使用具有N-末端截短缺失的某些糖基转移酶变体。 发现人β-半乳糖苷-α-2,6-唾液酸转移酶I（hST6Gal-1）的两种不同截短变体的组合表现出不同的特异性唾液酸转移酶酶活性。 在一个实例中，在第一变体Δ89hST6Gal-1催化双唾液酸化靶分子的形成的条件下，第二变体Δ108hST6Gal-1催化单唾液酸化靶分子的形成。 因此，公开了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，特别是以定量控制的方式唾液酸化作为糖蛋白部分的聚糖部分的末端受体基团，例如 免疫球蛋白。