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1.发明授权公开(公告)号：US08173416B2
公开(公告)日：2012-05-08
申请号：US10475840
申请日：2002-04-23
申请人： Herbert Müller-Hartmann , Gudula Riemen , Kirsten Rothmann-Cosic , Corinna Thiel , Ludger Altrogge , Meike Weigel , Rainer Christine , Elke Lorbach , Juliana Helfrich , Heike Wessendorf , Gregor Siebenkotten
发明人： Herbert Müller-Hartmann , Gudula Riemen , Kirsten Rothmann-Cosic , Corinna Thiel , Ludger Altrogge , Meike Weigel , Rainer Christine , Elke Lorbach , Juliana Helfrich , Heike Wessendorf , Gregor Siebenkotten
CPC分类号： A61N1/327 , A61N1/0412 , C12N15/87 , H03K3/57
摘要： The invention relates to a novel circuit arrangement for electrotransfection or electrofusion, which enables the transportation of DNA and/or other biologically active molecules to the nucleus of higher eukaryotic cells or the fusion of cells, independent of cell division and with reduced cell mortality.
摘要翻译： 本发明涉及一种用于电转移或电融合的新型电路装置，其能够将DNA和/或其他生物活性分子运输到高等真核细胞的细胞核或细胞融合，而与细胞分裂无关并具有降低的细胞死亡率。
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2.发明授权Buffer solution for electroporation and a method comprising the use of the same 有权用于电穿孔的缓冲溶液和包括其使用的方法公开(公告)号：US07332332B2
公开(公告)日：2008-02-19
申请号：US10475967
申请日：2002-04-23
申请人： Gudula Riemen , Elke Lorbach , Juliana Helfrich , Gregor Siebenkotten , Herbert Müller-Hartmann , Kirsten Rothmann , Corinna Thiel , Meike Weigel , Heike Wessendorf , Helmut Brosterbus , Michael Nix
发明人： Gudula Riemen , Elke Lorbach , Juliana Helfrich , Gregor Siebenkotten , Herbert Müller-Hartmann , Kirsten Rothmann , Corinna Thiel , Meike Weigel , Heike Wessendorf , Helmut Brosterbus , Michael Nix
摘要： The invention relates to a buffer solution for suspending animal or human cells and for dissolving biologically active molecules in order to introduce biologically active molecules into cells using an electric current. The inventive buffer solution has a buffering capacity of at least 20 mmol−1×pH−1 and an ionic strength of at least 200 mmol×1−1 during a change to the pH value from pH 7 to pH 8 and at a temperature of 25° C. The use of a buffer solution of this type allows biologically active molecules to be introduced into animal and human cells with a high degree of transfection efficiency and at the same time a low cell mortality. Different cell types, in particular dormant and actively dividing cells of low activity, can be successfully transfected in the buffer solution.
摘要翻译： 本发明涉及用于悬浮动物或人细胞并溶解生物活性分子以便使用电流将生物活性分子引入细胞的缓冲溶液。 本发明的缓冲溶液具有至少20mmol -1pp-1的缓冲能力，并且离子强度至少为200mmol / 在pH值从pH7到pH8改变到25℃的温度下。使用这种缓冲溶液使生物活性分子以高转染效率被引入动物和人细胞中 同时细胞死亡率低。 不同的细胞类型，特别是休眠和活跃分裂细胞的低活性，可以成功地转染到缓冲溶液中。
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3.发明申请Buffer solution for electroporation and a method comprising the use of the same 有权用于电穿孔的缓冲溶液和包括其使用的方法公开(公告)号：US20050064596A1
公开(公告)日：2005-03-24
申请号：US10475967
申请日：2002-04-23
申请人： Gudula Riemen , Elke Lorbach , Juliana Helfrich , Gregor Siebenkotten , Herbert Muller-Hartmann , Kirsten Rothmann-Cosic , Corinna Thiel , Meike Weigel , Heike Wessendorf , Helmut Brosterbus , Michael Nix
发明人： Gudula Riemen , Elke Lorbach , Juliana Helfrich , Gregor Siebenkotten , Herbert Muller-Hartmann , Kirsten Rothmann-Cosic , Corinna Thiel , Meike Weigel , Heike Wessendorf , Helmut Brosterbus , Michael Nix
IPC分类号： C12N15/09 , A61K35/12 , A61K38/00 , A61K47/00 , A61K48/00 , C12N13/00 , C12N15/87 , G01N33/50
摘要： The invention relates to a buffer solution for suspending animal or human cells and for dissolving biologically active molecules in order to introduce said biologically active molecules into the cells using an electric current and to a method for introducing biologically active molecules into animal or human cells using an electric current and a buffer solution. The inventive buffer solution has a buffering capacity of at least 20 mmol*I−1*pH−1 and an ionic strength of at least 200 mmol*I−1 during a change to the pH value from pH 7 to pH 8 and at a temperature of 25° C. The use of a buffer solution of this type in the corresponding method allows biologically active molecules to be introduced into animal and human cells with a high degree of transfection efficiency and at the same time a low cell mortality. Different cell types, in particular dormant and actively dividing cells of low activity, can be successfully transfected in said buffer solution.
摘要翻译： 本发明涉及一种用于悬浮动物或人细胞并用于溶解生物活性分子以便使用电流将所述生物活性分子引入细胞的缓冲溶液以及使用以下方法将生物活性分子引入动物或人细胞的方法： 电流和缓冲液。 本发明的缓冲溶液在pH值从pH7变化期间具有至少20mmol * I -1 * pH <-1的缓冲能力和至少200mmol * I -1的离子强度 至pH8，温度为25℃。在相应方法中使用这种类型的缓冲溶液使生物活性分子以高度的转染效率被引入动物和人类细胞，并且同时 细胞死亡率低。 可以在所述缓冲溶液中成功转染不同细胞类型，特别是低活性的休眠和活跃分化细胞。
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4.发明授权Buffer solution for electroporation and a method comprising the use of the same 有权用于电穿孔的缓冲溶液和包括其使用的方法公开(公告)号：US08192990B2
公开(公告)日：2012-06-05
申请号：US13009024
申请日：2011-01-19
申请人： Gudula Riemen , Elke Lorbach , Juliana Helfrich , Gregor Siebenkotten , Herbert Mueller-Hartmann , Kirsten Rothmann-Cosic , Corinna Thiel , Meike Weigel , Heike Wessendorf , Helmut Brosterhus , Michael Nix
发明人： Gudula Riemen , Elke Lorbach , Juliana Helfrich , Gregor Siebenkotten , Herbert Mueller-Hartmann , Kirsten Rothmann-Cosic , Corinna Thiel , Meike Weigel , Heike Wessendorf , Helmut Brosterhus , Michael Nix
摘要： A method for introducing biologically active molecules into animal or human cells using an electric current includes suspending the cells and dissolving the biologically active molecules in a buffer solution including HEPES and at least 10 mmol×1−1 magnesium ions (Mg2+), the buffer solution having a buffer capacity of at least 20 mmol×1−1 ×pH−1 at a change in the pH from pH 7 to pH 8 and at a temperature of 25° C., and an ionic strength of at least 200 mmol×1−1. An electric voltage is applied to the suspension.
摘要翻译： 使用电流将生物活性分子引入动物或人细胞的方法包括悬浮细胞并将生物活性分子溶解在包括HEPES和至少10mmol×1-1的镁离子（Mg 2+）的缓冲溶液中，缓冲溶液 在pH7至pH8和25℃的温度下，缓冲容量至少为20mmol×1-1×pH-1，离子强度为至少200mmol×1 -1。 向悬架施加电压。
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5.发明申请Differentially expressed tumour-specific polypeptides for use in the diagnosis and treatment of cancer 审中-公开用于诊断和治疗癌症的差异表达的肿瘤特异性多肽公开(公告)号：US20090252721A1
公开(公告)日：2009-10-08
申请号：US10571167
申请日：2004-09-20
申请人： Thomas Buschmann , Nikoleta-Kyriaki Fotiadis , Miriam Fuchs , Steffen Heim , Tatiana Lehnherr-Ilina , Stephanie Lamer , Jörn Meuer , Kirsten Rothmann-Cosic , Volker Seibert , Silvia Tortola Perez , Katrin Stedronsky
发明人： Thomas Buschmann , Nikoleta-Kyriaki Fotiadis , Miriam Fuchs , Steffen Heim , Tatiana Lehnherr-Ilina , Stephanie Lamer , Jörn Meuer , Kirsten Rothmann-Cosic , Volker Seibert , Silvia Tortola Perez , Katrin Stedronsky
IPC分类号： A61K39/395 , G01N33/566 , C40B30/04 , A61K31/7105 , A61K45/00 , C07K16/00 , C12Q1/68 , A61P35/00
CPC分类号： C12Q1/6886 , C12Q2600/106 , C12Q2600/136 , C12Q2600/158 , C12Q2600/178 , G01N33/57407 , G01N33/6893 , G01N2500/00
摘要： The invention relates to agents and methods for the diagnosis, prognosis and treatment of cancer. Specifically, the invention relates to the use of nucleic and amino acid sequences encoding transmembrane superfamily member 6 (TM4SF6), synaptophysin like protein (SYPL), stomatin like 2 (STOML2), Ras related GTP binding protein RAGA), nucleotide sensitive chloride channel 1A (CLNS1A), prion protein (p27-30) (PRNP), guanine nucleotide binding protein beta 2-like 1 (GNB2L1), guanine nucleotide binding protein 4 (GNG4), integral membrane protein 2B (ITM2B), integral membrane protein 1 (ITM1), transmembrane 9 superfamily member 2 (TM9SF2), opiate receptor-like 1 protein (OPRL1), low density lipoprotein receptor-related protein 4 (LRP4), human glomerular epithelial protein 1 (GLEPP1), toll-like receptor 3 (TLR3), and/or zona pellucida glycoprotein 3A (ZP3) for the diagnosis of both early and late stage non-steroid specific cancers, cancer prognosis, as well as screening for therapeutic agents that regulate the gene expression and/or biological activity of said proteins. This invention further relates to the biological technologies designed to inhibit the gene expression and/or biological activity of said proteins including using agents identified in screening assays described herein, vector delivery of antisense polynucleotide sequences, and antibody targeting of said proteins. In specific embodiments, the proteins are of human origin.
摘要翻译： 本发明涉及用于诊断，预后和治疗癌症的药剂和方法。 具体而言，本发明涉及编码跨膜超家族成员6（TM4SF6），突触素样蛋白（SYPL），类似2的孔蛋白（STOML2），Ras相关GTP结合蛋白RAGA的核酸和氨基酸序列，核苷酸敏感性氯通道1A （CLNS1A），朊病毒蛋白（p27-30）（PRNP），鸟嘌呤核苷酸结合蛋白β2-样1（GNB2L1），鸟嘌呤核苷酸结合蛋白4（GNG4），整合膜蛋白2B（ITM2B），整合膜蛋白1 ITM1），跨膜9超家族成员2（TM9SF2），阿片受体样1蛋白（OPRL1），低密度脂蛋白受体相关蛋白4（LRP4），人肾小球上皮蛋白1（GLEPP1），toll样受体3（TLR3 ）和/或透明带糖蛋白3A（ZP3），用于诊断早期和晚期非类固醇特异性癌症，癌症预后，以及筛选调节所述蛋白质的基因表达和/或生物活性的治疗剂 。 本发明还涉及设计用于抑制所述蛋白质的基因表达和/或生物活性的生物学技术，包括使用本文所述的筛选测定中鉴定的试剂，反义多核苷酸序列的载体递送和所述蛋白质的抗体靶向。 在具体实施方案中，蛋白质是人源的。
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6.发明授权Buffer solution for electroporation and a method comprising the use of the same 有权用于电穿孔的缓冲溶液和包括其使用的方法公开(公告)号：US08039259B2
公开(公告)日：2011-10-18
申请号：US11931996
申请日：2007-10-31
申请人： Gudula Riemen , Elke Lorbach , Juliana Helfrich , Gregor Siebenkotten , Herbert Mueller-Hartmann , Kirsten Rothmann-Cosic , Corinna Thiel , Meike Weigel , Heike Wessendorf , Helmut Brosterhus , Michael Nix
发明人： Gudula Riemen , Elke Lorbach , Juliana Helfrich , Gregor Siebenkotten , Herbert Mueller-Hartmann , Kirsten Rothmann-Cosic , Corinna Thiel , Meike Weigel , Heike Wessendorf , Helmut Brosterhus , Michael Nix
摘要： A buffer solution for suspending animal or human cells and for dissolving biologically active molecules in order to introduce the biologically active molecules into the cells using electric current. The buffer solution includes at least one of sodium succinate, mannitol and sodium lactobionate. The buffer solution has a buffer capacity of at least 20 mmol*l−1*pH−1 at a change in the from pH 7 to pH 8 and at a temperature of 25° C., and an ionic strength of at least 200 mmol * l−1.
摘要翻译： 用于悬浮动物或人细胞并溶解生物活性分子以便使用电流将生物活性分子引入细胞的缓冲溶液。 缓冲溶液包括琥珀酸钠，甘露糖醇和乳糖酸钠中的至少一种。 缓冲溶液在pH7至pH8的变化和25℃的温度下具有至少20mmol * l-1 * pH-1的缓冲容量，离子强度至少为200mmol * l-1。
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7.发明申请BUFFER SOLUTION FOR ELECTROPORATION AND A METHOD COMPRISING THE USE OF THE SAME 有权用于电镀的缓冲溶液和包含其使用的方法公开(公告)号：US20110117654A1
公开(公告)日：2011-05-19
申请号：US13009024
申请日：2011-01-19
申请人： Gudula Riemen , Elke Lorbach , Juliana Helfrich , Gregor Siebenkotten , Herbert Mueller-Hartmann , Kirsten Rothmann-Cosic , Corinna Thiel , Meike Weigel , Heike Wessendorf , Helmut Brosterhaus , Michael Nix
发明人： Gudula Riemen , Elke Lorbach , Juliana Helfrich , Gregor Siebenkotten , Herbert Mueller-Hartmann , Kirsten Rothmann-Cosic , Corinna Thiel , Meike Weigel , Heike Wessendorf , Helmut Brosterhaus , Michael Nix
摘要： A method for introducing biologically active molecules into animal or human cells using an electric current includes suspending the cells and dissolving the biologically active molecules in a buffer solution including HEPES and at least 10 mmol×1−1 magnesium ions (Mg2+), the buffer solution having a buffer capacity of at least 20 mmol×1−1×pH−1 at a change in the pH from pH 7 to pH 8 and at a temperature of 25° C., and an ionic strength of at least 200 mmol×1−1. An electric voltage is applied to the suspension.
摘要翻译： 使用电流将生物活性分子引入动物或人细胞的方法包括悬浮细胞并将生物活性分子溶解在包括HEPES和至少10mmol×1-1的镁离子（Mg 2+）的缓冲溶液中，缓冲溶液 在pH7至pH8和25℃的温度下，缓冲容量至少为20mmol×1-1×pH-1，离子强度为至少200mmol×1 -1。 向悬架施加电压。
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8.发明申请公开(公告)号：US20090023131A1
公开(公告)日：2009-01-22
申请号：US11836656
申请日：2007-08-09
申请人： Herbert Muller-Hartmann , Gudula Riemen , Kirsten Rothmann-Cosic , Corinna Thiel , Ludger Altrogge , Meike Weigel , Rainer Christine , Elke Lorbach , Juliana Helfrich , Heike Wessendorf , Gregor Siebenkotten
发明人： Herbert Muller-Hartmann , Gudula Riemen , Kirsten Rothmann-Cosic , Corinna Thiel , Ludger Altrogge , Meike Weigel , Rainer Christine , Elke Lorbach , Juliana Helfrich , Heike Wessendorf , Gregor Siebenkotten
IPC分类号： C12Q3/00
CPC分类号： A61N1/327 , A61N1/0412 , C12N15/87 , H03K3/57
摘要： The invention relates to a novel circuit arrangement for electrotransfection or electrofusion, which enables the transportation of DNA and/or other biologically active molecules to the nucleus of higher eukaryotic cells or the fusion of cells, independent of cell division and with reduced cell mortality.
摘要翻译： 本发明涉及一种用于电转移或电融合的新型电路装置，其能够将DNA和/或其他生物活性分子运输到高等真核细胞的细胞核或细胞融合，而与细胞分裂无关并具有降低的细胞死亡率。
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9.发明申请Buffer solution for electroporation and a method comprising the use of the same 有权用于电穿孔的缓冲溶液和包括其使用的方法公开(公告)号：US20070110736A1
公开(公告)日：2007-05-17
申请号：US11595448
申请日：2006-11-09
申请人： Gudula Riemen , Elke Lorbach , Juliana Helfrich , Gregor Siebenkotten , Herbert Muller-Hartmann , Kirsten Rothmann-Cosic , Corinna Thiel , Meike Weigel , Heike Wessendorf , Helmut Brosterhus , Michael Nix
发明人： Gudula Riemen , Elke Lorbach , Juliana Helfrich , Gregor Siebenkotten , Herbert Muller-Hartmann , Kirsten Rothmann-Cosic , Corinna Thiel , Meike Weigel , Heike Wessendorf , Helmut Brosterhus , Michael Nix
摘要： The invention relates to a buffer solution for suspending animal or human cells and for dissolving biologically active molecules in order to introduce said biologically active molecules into the cells using an electric current and to a method for introducing biologically active molecules into animal or human cells using an electric current and a buffer solution. The inventive buffer solution has a buffering capacity of at least 20 mmol*I−1*pH−1 and an ionic strength of at least 200 mmol* I−1 during a change to the pH value from pH 7 to pH 8 and at a temperature of 25 ° C. The use of a buffer solution of this type in the corresponding method allows biologically active molecules to be introduced into animal and human cells with a high degree of transfection efficiency and at the same time a low cell mortality. Different cell types, in particular dormant and actively dividing cells of low activity, can be successfully transfected in said buffer solution.
摘要翻译： 本发明涉及一种用于悬浮动物或人细胞并用于溶解生物活性分子以便使用电流将所述生物活性分子引入细胞的缓冲溶液以及使用以下方法将生物活性分子引入动物或人细胞的方法： 电流和缓冲液。 本发明缓冲溶液的缓冲能力至少为20毫摩尔/升，pH为-1，离子强度为至少200毫摩尔/升， 在pH值从pH7变化到pH8至25℃的温度下，在1℃下使用。在相应方法中使用这种类型的缓冲溶液允许将生物活性分子引入动物和 人细胞具有高转染效率，同时细胞死亡率低。 可以在所述缓冲溶液中成功转染不同细胞类型，特别是低活性的休眠和活跃分化细胞。
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10.发明授权Buffer solution for electroporation and a method comprising the use of the same 有权用于电穿孔的缓冲溶液和包括其使用的方法公开(公告)号：US08003389B2
公开(公告)日：2011-08-23
申请号：US11595448
申请日：2006-11-09
申请人： Gudula Riemen , Elke Lorbach , Juliana Helfrich , Gregor Siebenkotten , Herbert Mueller-Hartmann , Kirsten Rothmann-Cosic , Corinna Thiel , Meike Weigel , Heike Wessendorf , Helmut Brosterhus , Michael Nix
发明人： Gudula Riemen , Elke Lorbach , Juliana Helfrich , Gregor Siebenkotten , Herbert Mueller-Hartmann , Kirsten Rothmann-Cosic , Corinna Thiel , Meike Weigel , Heike Wessendorf , Helmut Brosterhus , Michael Nix
摘要： A method for introducing biologically active molecules into animal or human cells using electric current includes suspending the cells and dissolving the biologically active molecules in a buffer solution which has a buffer capacity of at least 20 mmol×l−1×pH−1 and an ionic strength of at least 200 mmol×l−1 at a change in the pH from pH 7 to pH 8 and at a temperature of 25° C. to form a suspension. The method further includes applying an electric voltage to the suspension so as to introduce the biologically active molecules into animal or human cells.
摘要翻译： 使用电流将生物活性分子引入动物或人细胞的方法包括悬浮细胞并将生物活性分子溶解在缓冲溶液中，缓冲溶液的缓冲容量至少为20mmol×l-1×pH-1，离子 强度为至少200mmol / l-1，pH在pH7至pH8和25℃温度下形成悬浮液。 该方法还包括向悬浮液施加电压以将生物活性分子引入动物或人细胞中。
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