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公开(公告)号：US20150184138A1
公开(公告)日：2015-07-02
申请号：US14448545
申请日：2014-07-31
申请人： Greg Pogue , Ernie Hiatt , Romy Kandzia , Stefan Werner , Frank Thieme , Tsafrir Mor
发明人： Greg Pogue , Ernie Hiatt , Romy Kandzia , Stefan Werner , Frank Thieme , Tsafrir Mor
CPC分类号： C12N9/18 , A61K38/465 , C12Y301/01008
摘要： A new, reliable, easily scalable and reproducible method for the production of recombinant butyrylcholinesterase (rBuChE) is provided. Through the utilization of a plant transfection procedure, various plant strains have been shown to generate effective and scalable amounts of rBuChE under acceptable manufacturing processes to permit reliable levels of such enzymes for desired nerve agent protection requirements (including tetrameric products). As well, such methods in engineered plant lines have shown suitable production of these enzymes in tetramer form with glycan formation and sialyalation (for terminal groups) to allow for optimal potency against organophosphorus agent exposure as well as proper immunogenic response within the plant sources. The overall production method, including the transfection and production within mammalian cells, as well as the process steps involved for such a reliable sourcing platform from plants is thus encompassed within the invention.
摘要翻译： 提供了一种用于生产重组丁酰胆碱酯酶（rBuChE）的新的，可靠的，易于扩展和重现的方法。 通过利用植物转染方法，已经显示各种植物菌株在可接受的制造过程中产生有效和可缩放量的rBuChE，以允许可靠水平的这些酶用于所需的神经药物保护要求（包括四聚物产品）。 同样，工程化植物品系中的这些方法已经显示出以四聚体形式合适地生产这些酶，其中聚糖形成和sialyalation（用于末端基团）以允许针对有机磷剂暴露的最佳效力以及在植物来源内的适当的免疫原性反应。 因此，包括哺乳动物细胞内的转染和生产的总体生产方法以及从植物获得这种可靠的采购平台所涉及的工艺步骤。
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公开(公告)号：US08883420B2
公开(公告)日：2014-11-11
申请号：US13121019
申请日：2009-10-08
申请人： Sylvestre Marillonnet , Stefan Werner , Carola Engler , Romy Kandzia , Frank Thieme , Ernst Weber
发明人： Sylvestre Marillonnet , Stefan Werner , Carola Engler , Romy Kandzia , Frank Thieme , Ernst Weber
摘要： A process of inserting a nucleic acid sequence of interest into an acceptor nucleic acid is provided. The process comprises amplifying by PCR a DNA comprising in the following order a sequence segment U, a nucleic acid sequence segment of known nucleotide sequence K2, and a nucleic acid sequence segment of known sequence K3. The process further comprises treating the linear double-stranded DNA molecules from the PCR amplification with an exonuclease to obtain a single-stranded overhang at the first end of the DNA and a single-stranded overhang comprising nucleic acid segments K2 and K3 at the second end of the DNA. The process additionally comprises annealing the product of the exonuclease treatment to a linearized double-stranded acceptor nucleic acid which has been designed to complement the single-stranded overhangs of the product of the exonuclease treatment.
摘要翻译： 提供将感兴趣的核酸序列插入受体核酸的过程。 该方法包括通过PCR扩增包含以下顺序的序列片段U，已知核苷酸序列K2的核酸序列片段和已知序列K3的核酸序列片段的DNA。 该方法还包括用外切核酸酶从PCR扩增中处理线性双链DNA分子，以在DNA的第一端获得单链突出端，并且在第二末端包含包含核酸片段K2和K3的单链突出端 的DNA。 该方法还包括将外切核酸酶处理的产物退火至线性化双链受体核酸，其被设计为补充外切核酸酶处理产物的单链突出端。
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公开(公告)号：US20110263024A1
公开(公告)日：2011-10-27
申请号：US13121019
申请日：2009-10-08
申请人： Sylvestre Marillonnet , Stefan Werner , Carola Engler , Romy Kandzia , Frank Thieme , Ernst Weber
发明人： Sylvestre Marillonnet , Stefan Werner , Carola Engler , Romy Kandzia , Frank Thieme , Ernst Weber
IPC分类号： C12N15/87
摘要： A process of inserting a nucleic acid sequence of interest into an acceptor nucleic acid, comprising the following steps: amplifying by PCR a DNA comprising in the following order a sequence segment U, a nucleic acid sequence segment of known nucleotide sequence K2 and a nucleic acid sequence segment of known sequence K3 using a forward primer defining a first end of the amplified DNA and a reverse primer defining a second end of the amplified DNA, said reverse primer terminating at its 3′-end in a nucleotide sequence of nucleic acid sequence segment K3; treating the linear double-stranded DNA molecules contained in the PCR product obtained in the previous step with an exonuclease to obtain a single-stranded overhang at the first end of the DNA and a single-stranded overhang comprising nucleic acid segments K2 and K3 at the second end of the DNA; and annealing the product of the previous step to a linearized double-stranded acceptor nucleic acid having at a first end thereof a single-stranded overhang complementary to the single-stranded overhang of the first end of the DNA and at a second end thereof a single-stranded overhang complementary to the single-stranded sequence segment K2 of the second end of the DNA.
摘要翻译： 包括以下步骤：通过PCR扩增包含以下顺序的DNA序列的已知核苷酸序列K2的核酸序列片段和核酸的方法 已知序列K3的序列片段使用限定扩增DNA第一末端的正向引物和限定扩增DNA第二末端的反向引物，所述反向引物在其核酸序列片段的核苷酸序列的3'末端终止 K3; 用外切核酸酶处理前一步骤中获得的PCR产物中包含的线性双链DNA分子，以在DNA的第一端获得单链突出端，并且在第一端包含核酸片段K2和K3的单链突出端 DNA的第二端; 并将前述步骤的产物退火到线性双链受体核酸，其在第一端具有与DNA的第一末端的单链突出部互补的单链突出端，并且在其第二端具有单链突出端 与DNA的第二末端的单链序列片段K2互补的突出突出端。
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公开(公告)号：US20170313999A1
公开(公告)日：2017-11-02
申请号：US15602622
申请日：2017-05-23
发明人： Greg Pogue , Ernie Hiatt , Romy Kandzia , Stefan Werner , Frank Thieme , Tsafrir Mor , Steven Hume
CPC分类号： C12N9/18 , A61K38/465 , C12Y301/01008
摘要： A new, reliable, easily scalable and reproducible method for the production of recombinant butyrylcholinesterase (rBuChE) is provided. Through the utilization of a plant transfection procedure, various plant strains have been shown to generate effective and scalable amounts of rBuChE under acceptable manufacturing processes to permit reliable levels of such enzymes for desired nerve agent protection requirements (including tetrameric products). As well, such methods in engineered plant lines have shown suitable production of these enzymes in tetramer form with glycan formation and sialylation (for terminal groups) to allow for optimal potency against organophosphorus agent exposure as well as proper immunogenic response within the plant sources. The overall production method, including the transfection and production within mammalian cells, as well as the process steps involved for such a reliable sourcing platform from plants is thus encompassed within the invention.
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公开(公告)号：US10035994B2
公开(公告)日：2018-07-31
申请号：US15602622
申请日：2017-05-23
发明人： Greg Pogue , Ernie Hiatt , Romy Kandzia , Stefan Werner , Frank Thieme , Tsafrir Mor , Steven Hume
CPC分类号： C12N9/18 , A61K38/465 , C12Y301/01008
摘要： A new, reliable, easily scalable and reproducible method for the production of recombinant butyrylcholinesterase (rBuChE) is provided. Through the utilization of a plant transfection procedure, various plant strains have been shown to generate effective and scalable amounts of rBuChE under acceptable manufacturing processes to permit reliable levels of such enzymes for desired nerve agent protection requirements (including tetrameric products). As well, such methods in engineered plant lines have shown suitable production of these enzymes in tetramer form with glycan formation and sialylation (for terminal groups) to allow for optimal potency against organophosphorus agent exposure as well as proper immunogenic response within the plant sources. The overall production method, including the transfection and production within mammalian cells, as well as the process steps involved for such a reliable sourcing platform from plants is thus encompassed within the invention.
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公开(公告)号：US09688970B2
公开(公告)日：2017-06-27
申请号：US14448545
申请日：2014-07-31
发明人： Greg Pogue , Ernie Hiatt , Romy Kandzia , Stefan Werner , Frank Thieme , Tsafrir Mor , Steven Hume
CPC分类号： C12N9/18 , A61K38/465 , C12Y301/01008
摘要： A new, reliable, easily scalable and reproducible method for the production of recombinant butyrylcholinesterase (rBuChE) is provided. Through the utilization of a plant transfection procedure, various plant strains have been shown to generate effective and scalable amounts of rBuChE under acceptable manufacturing processes to permit reliable levels of such enzymes for desired nerve agent protection requirements (including tetrameric products). As well, such methods in engineered plant lines have shown suitable production of these enzymes in tetramer form with glycan formation and sialyalation (for terminal groups) to allow for optimal potency against organophosphorus agent exposure as well as proper immunogenic response within the plant sources. The overall production method, including the transfection and production within mammalian cells, as well as the process steps involved for such a reliable sourcing platform from plants is thus encompassed within the invention.
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公开(公告)号：US07969139B2
公开(公告)日：2011-06-28
申请号：US12519812
申请日：2007-12-12
申请人： Jochen Ermisch , Uwe Hering , Frank Thieme , Ralf-Reiner Volkmar
发明人： Jochen Ermisch , Uwe Hering , Frank Thieme , Ralf-Reiner Volkmar
IPC分类号： G01R33/00
CPC分类号： G01R15/181
摘要： A Rogowski sensor is provided for measuring a current of an electrical conductor. The sensor has a Rogowski coil, which has an electrical winding resistor, and an integration device which is connected to the Rogowski coil and is intended to generate an output signal which is proportional to the electrical current to be measured by the Rogowski sensor. The integration device is formed by the winding resistor of the Rogowski coil and a capacitance which is connected to the two winding connections of the Rogowski coil, and for the output signal from the integration device to be formed by the voltage applied to the capacitance.
摘要翻译： 提供Rogowski传感器用于测量电导体的电流。 传感器具有罗氏线圈，其具有电绕组电阻器以及与罗氏线圈连接的集成装置，用于产生与由罗戈夫斯基传感器测量的电流成比例的输出信号。 集成装置由Rogowski线圈的绕组电阻器和连接到Rogowski线圈的两个绕组连接的电容器组成，并且来自积分器件的输出信号由施加到电容器的电压形成。
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公开(公告)号：US06346805B1
公开(公告)日：2002-02-12
申请号：US09291665
申请日：1999-04-14
申请人： Jochen Ermisch , Uwe Stephan , Frank Thieme , Peter Bauerschmidt
发明人： Jochen Ermisch , Uwe Stephan , Frank Thieme , Peter Bauerschmidt
IPC分类号： G01R1900
CPC分类号： G01R15/146
摘要： For precise measurement of currents with high dynamic range, especially in gas-insulated voltage switchgear, a first signal of a first low-power measurement transformer and a second signal of a second nonmagnetic measurement transformer, operating, for example, according to the principle of a Rogowski coil or according to the Faraday effect, are compared. Depending on the degree of accuracy, one of the two signals is then selected and further processed. A deviation of the two signals from each other, especially their gradient, is referred to as comparison criterion. In a corresponding configuration, the two measurement transformers of different type are connected to a common processing component for signal processing.
摘要翻译： 对于具有高动态范围的电流的精确测量，特别是在气体绝缘电压开关设备中，第一低功率测量变压器的第一信号和第二非磁性测量变压器的第二信号，例如根据 罗格斯基线圈或根据法拉第效应进行比较。 根据准确度，然后选择两个信号之一并进一步处理。 两个信号彼此的偏差，特别是它们的梯度，被称为比较标准。 在相应的配置中，不同类型的两个测量变压器连接到用于信号处理的公共处理部件。
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公开(公告)号：US20100013460A1
公开(公告)日：2010-01-21
申请号：US12519812
申请日：2007-12-12
申请人： Jochen Ermisch , Uwe Hering , Frank Thieme , Ralf-Reiner Volkmar
发明人： Jochen Ermisch , Uwe Hering , Frank Thieme , Ralf-Reiner Volkmar
IPC分类号： G01R15/18
CPC分类号： G01R15/181
摘要： A Rogowski sensor is provided for measuring a current of an electrical conductor. The sensor has a Rogowski coil, which has an electrical winding resistor, and an integration device which is connected to the Rogowski coil and is intended to generate an output signal which is proportional to the electrical current to be measured by the Rogowski sensor. The integration device is formed by the winding resistor of the Rogowski coil and a capacitance which is connected to the two winding connections of the Rogowski coil, and for the output signal from the integration device to be formed by the voltage applied to the capacitance.
摘要翻译： 提供Rogowski传感器用于测量电导体的电流。 传感器具有罗氏线圈，其具有电绕组电阻器以及与罗氏线圈连接的集成装置，用于产生与由罗戈夫斯基传感器测量的电流成比例的输出信号。 集成装置由Rogowski线圈的绕组电阻器和连接到Rogowski线圈的两个绕组连接的电容器组成，并且来自积分器件的输出信号由施加到电容器的电压形成。
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