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公开(公告)号：US09574186B2
公开(公告)日：2017-02-21
申请号：US14697843
申请日：2015-04-28
Applicant: Roche Diagnostics Operations, Inc.
Inventor： Werner Hoelke , Johann-Peter Thalhofer , Antje Liehre , Markus Weber
CPC classification number: C12N9/54 , C12N9/96 , C12Y304/24027
Abstract: The present invention deals with the proteolytic enzyme thermolysin which tends to be unstable in aqueous solution. The invention provides methods and compositions to enhance the stability of dissolved thermolysin in aqueous solution. Thermolysin, crude thermolysin or a lyophilisate containing thermolysin and one or more salts, is contacted with an aqueous buffer with a low salt concentration and a first solution is formed. Subsequently, a further salt in solid form is added and dissociated, thereby forming a second solution comprising thermolysin in a stabilized form.
Abstract translation: 本发明涉及在水溶液中趋于不稳定的蛋白水解酶嗜热溶菌酶。 本发明提供了增加溶解的嗜热溶菌酶在水溶液中的稳定性的方法和组合物。 将含有嗜热菌蛋白酶和一种或多种盐的溶血素酶，粗嗜热菌蛋白酶或冻干物质与低盐浓度的水性缓冲液接触，形成第一溶液。 随后，加入固体形式的另外的盐并解离，由此形成稳定形式的包含嗜热菌蛋白酶的第二溶液。
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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.
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公开(公告)号：US20150337282A1
公开(公告)日：2015-11-26
申请号：US14697843
申请日：2015-04-28
Applicant: Roche Diagnostics Operations, Inc.
Inventor： Werner Hoelke , Johann-Peter Thalhofer , Antje Liehre , Markus Weber
IPC: C12N9/54
CPC classification number: C12N9/54 , C12N9/96 , C12Y304/24027
Abstract: The present invention deals with the proteolytic enzyme thermolysin which tends to be unstable in aqueous solution. The invention provides methods and compositions to enhance the stability of dissolved thermolysin in aqueous solution. Thermolysin, crude thermolysin or a lyophilisate containing thermolysin and one or more salts, is contacted with an aqueous buffer with a low salt concentration and a first solution is formed. Subsequently, a further salt in solid form is added and dissociated, thereby forming a second solution comprising thermolysin in a stabilized form.
Abstract translation: 本发明涉及在水溶液中趋于不稳定的蛋白水解酶嗜热溶菌酶。 本发明提供了增加溶解的嗜热溶菌酶在水溶液中的稳定性的方法和组合物。 将含有嗜热菌蛋白酶和一种或多种盐的溶血素酶，粗嗜热菌蛋白酶或冻干物质与低盐浓度的水性缓冲液接触，形成第一溶液。 随后，加入固体形式的另外的盐并解离，由此形成稳定形式的包含嗜热菌蛋白酶的第二溶液。
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公开(公告)号：US20230052545A1
公开(公告)日：2023-02-16
申请号：US17822587
申请日：2022-08-26
Applicant: Roche Diagnostics Operations, Inc.
Inventor： Werner Hoelke , Michaela Fischer , Johann-Peter Thalhofer , Markus Weber
Abstract: Described are compositions, in particular lyophilizates, containing proteolytic enzymes, and methods for producing the compositions. Typically these compositions contain one or more proteases with collagenase activity and a neutral protease, for example, thermolysin. The compositions are free of acetate salts. Surprisingly, such compositions can be dissolved in water more rapidly than lyophilized protease mixtures of the state of the art.
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公开(公告)号：US20170247679A1
公开(公告)日：2017-08-31
申请号：US15595126
申请日：2017-05-15
Applicant: Roche Diagnostics Operations, Inc.
Inventor： Werner Hoelke , Michaela Fischer , Johann-Peter Thalhofer , Markus Weber
CPC classification number: C12N9/96 , C12N9/52 , C12N9/54 , C12N9/6491 , C12Y304/24003 , C12Y304/24027
Abstract: Described are compositions, in particular lyophilizates, containing proteolytic enzymes, and methods for producing the compositions. Typically these compositions contain one or more proteases with collagenase activity and a neutral protease, for example, thermolysin. The compositions are free of acetate salts. Surprisingly, such compositions can be dissolved in water more rapidly than lyophilized protease mixtures of the state of the art.
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Patent Agency Ranking
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。
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公开(公告)号：US11459556B2
公开(公告)日：2022-10-04
申请号：US16865487
申请日：2020-05-04
Applicant: Roche Diagnostics Operations, Inc.
Inventor： Werner Hoelke , Michaela Fischer , Johann-Peter Thalhofer , Markus Weber
Abstract: Described are compositions, in particular lyophilizates, containing proteolytic enzymes, and methods for producing the compositions. Typically these compositions contain one or more proteases with collagenase activity and a neutral protease, for example, thermolysin. The compositions are free of acetate salts. Surprisingly, such compositions can be dissolved in water more rapidly than lyophilized protease mixtures of the state of the art.
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公开(公告)号：US10227642B2
公开(公告)日：2019-03-12
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
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.
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公开(公告)号：US09663769B2
公开(公告)日：2017-05-30
申请号：US14872392
申请日：2015-10-01
Applicant: Roche Diagnostics Operations, Inc.
Inventor： Michael Greif , Christian Rudolph , Manfred Schmidt , Harald Sobek , Johann-Peter Thalhofer
IPC: C12N9/12
CPC classification number: C12N9/1247 , C12Y207/07006
Abstract: The present disclosure provide novel variants of T7 RNA polymerase. Embodiments of T7 variants, according to the instant invention, include a Cysteine-Serine substitution on position 723 of the amino acid sequence of the T7 polypeptide. Embodiments of T7 variants according to the instant invention have a DNA-dependent RNA polymerase enzymatic activity and a reduced tendency to form intramolecular homodimers by way of oxidizing thiol groups. The amino acid substitutions within the T7 variants disclosed herein impact minimally, if at all, the RNA polymerase activity of the T7 polypeptide. Further, the mutations of the disclosed embodiments may optionally be combined with mutations which provide enhanced thermostability compared to the wild-type reference.
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公开(公告)号：US20160032261A1
公开(公告)日：2016-02-04
申请号：US14885732
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
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.
Abstract translation: 本发明通过引入导致酶的热稳定性的新突变来提供T7RNA聚合酶的改进变体。 根据本发明，位置Val426，Ser633，Val650，Thr654，Ala702，Val795及其组合的氨基酸取代是有利的。
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