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51.发明授权Luminescence resonance energy transfer (LRET) assays for clostridial toxin activity 有权梭菌毒素活性的发光共振能量转移（LRET）测定公开(公告)号：US08022172B2
公开(公告)日：2011-09-20
申请号：US11752596
申请日：2007-05-23
申请人： Dudley J. Williams , Marcella A. Gilmore , Lance E. Steward , Marc Verhagen , Kei Roger Aoki , Ester Fernandez-Salas
发明人： Dudley J. Williams , Marcella A. Gilmore , Lance E. Steward , Marc Verhagen , Kei Roger Aoki , Ester Fernandez-Salas
IPC分类号： A61K38/00 , C07K7/00 , C07K17/00 , C07K14/00 , C07K16/00 , C07K5/00 , C12N1/20 , C12N11/00 , G01N33/533 , G01N33/554 , G01N33/569
CPC分类号： C12Q1/37 , C07K14/001 , C07K14/435 , G01N33/542 , G01N33/56911 , G01N2333/33
摘要： Clostridial toxin substrates comprising a lanthanide donor complex, an acceptor, and a Clostridial toxin recognition sequence including a cleavage site; methods for determining the activity of a Clostridial toxin from a test sample using such Clostridial toxin substrates; cell compositions comprising such Clostridial toxin substrates and a Clostridial toxin receptor; and methods for determining the activity of a Clostridial toxin from a test sample using such cell compositions.
摘要翻译： 包含镧系元素供体复合物，受体和包含切割位点的梭菌毒素识别序列的梭菌毒素底物; 使用这种梭菌毒素底物从测试样品测定梭菌毒素的活性的方法; 包含这种梭菌毒素底物和梭菌毒素受体的细胞组合物; 以及使用这种细胞组合物从测试样品测定梭菌毒素的活性的方法。
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52.发明授权Cell compositions useful for cell-based FRET assays for clostridial toxins 有权用于梭菌毒素的基于细胞的FRET测定的细胞组合物公开(公告)号：US07749759B2
公开(公告)日：2010-07-06
申请号：US12192925
申请日：2008-08-15
IPC分类号： C12N5/00
CPC分类号： C07K14/005 , C07K14/33 , C07K14/4702 , C07K14/4725 , C12N2710/16622 , C12N2740/16122 , C12Q1/37
摘要： The present invention provides a method of determining clostridial toxin activity by (a) contacting with a sample a cell containing a clostridial toxin substrate that includes a donor fluorophore; an acceptor having an absorbance spectrum overlapping the emission spectrum of the donor fluorophore; and a clostridial toxin recognition sequence containing a cleavage site that intervenes between the donor fluorophore and the acceptor, where resonance energy transfer is exhibited between the donor fluorophore and the acceptor under the appropriate conditions; (b) exciting the donor fluorophore; and (c) determining resonance energy transfer of the contacted cell relative to a control cell, where a difference in resonance energy transfer of the contacted cell as compared to the control cell is indicative of clostridial toxin activity.
摘要翻译： 本发明提供了一种通过（a）使样品与包含供体荧光团的梭菌毒素底物的细胞接触来测定梭菌毒素活性的方法; 具有与供体荧光团的发射光谱重叠的吸收光谱的受体; 以及包含介入供体荧光团和受体之间的切割位点的梭菌毒素识别序列，其中在适当条件下在供体荧光团和受体之间显示共振能量转移; （b）激发供体荧光团; 和（c）确定接触细胞相对于对照细胞的共振能量转移，其中与对照细胞相比，所接触的细胞的共振能量转移的差异指示梭菌毒素活性。
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公开(公告)号：US07718766B2
公开(公告)日：2010-05-18
申请号：US12042458
申请日：2008-10-31
CPC分类号： G01N33/582
摘要： The present invention provides clostridial toxin substrates useful in assaying for the protease activity of any clostridial toxin, including botulinum toxins of all serotypes as well as tetanus toxins. A clostridial toxin substrate of the invention contains a donor fluorophore; an acceptor having an absorbance spectrum overlapping the emission spectrum of the donor fluorophore; and a clostridial toxin recognition sequence that includes a cleavage site, where the cleavage site intervenes between the donor fluorophore and the acceptor and where, under the appropriate conditions, resonance energy transfer is exhibited between the donor fluorophore and the acceptor.
摘要翻译： 本发明提供了可用于测定任何梭菌毒素（包括所有血清型的肉毒毒素）以及破伤风毒素的蛋白酶活性的梭菌毒素底物。 本发明的梭菌毒素底物含有供体荧光团; 具有与供体荧光团的发射光谱重叠的吸收光谱的受体; 和梭菌毒素识别序列，其包括切割位点，其中切割位点插入供体荧光团和受体之间，并且在适当条件下，在供体荧光团和受体之间显示共振能量转移。
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54.发明申请FRET PROTEASE ASSAYS FOR BOTULINUM SEROTYPE A/E TOXINS 失效用于BOTULINUM SEROTYPE A / E毒素的FRET PROTEASE测定公开(公告)号：US20080146786A1
公开(公告)日：2008-06-19
申请号：US11609189
申请日：2006-12-11
IPC分类号： C07K14/195
CPC分类号： G01N33/582
摘要： The present invention provides clostridial toxin substrates useful in assaying for the protease activity of any clostridial toxin, including botulinum toxins of all serotypes as well as tetanus toxins. A clostridial toxin substrate of the invention contains a donor fluorophore; an acceptor having an absorbance spectrum overlapping the emission spectrum of the donor fluorophore; and a clostridial toxin recognition sequence that includes a cleavage site, where the cleavage site intervenes between the donor fluorophore and the acceptor and where, under the appropriate conditions, resonance energy transfer is exhibited between the donor fluorophore and the acceptor.
摘要翻译： 本发明提供了可用于测定任何梭菌毒素（包括所有血清型的肉毒毒素）以及破伤风毒素的蛋白酶活性的梭菌毒素底物。 本发明的梭菌毒素底物含有供体荧光团; 具有与供体荧光团的发射光谱重叠的吸收光谱的受体; 和梭菌毒素识别序列，其包括切割位点，其中切割位点插入供体荧光团和受体之间，并且在适当条件下，在供体荧光团和受体之间显示共振能量转移。
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55.发明授权Cell-based fluorescence resonance energy transfer (FRET) assays for clostridial toxins 有权梭菌毒素的基于细胞的荧光共振能量转移（FRET）测定公开(公告)号：US07183066B2
公开(公告)日：2007-02-27
申请号：US10261161
申请日：2002-09-27
IPC分类号： C12Q1/37 , C07H21/04 , C12P21/06 , C12N15/74 , C07N14/435
CPC分类号： C07K14/005 , C07K14/33 , C07K14/4702 , C07K14/4725 , C12N2710/16622 , C12N2740/16122 , C12Q1/37
摘要： The present invention provides a method of determining clostridial toxin activity by (a) contacting with a sample a cell containing a clostridial toxin substrate that includes a donor fluorophore; an acceptor having an absorbance spectrum overlapping the emission spectrum of the donor fluorophore; and a clostridial toxin recognition sequence containing a cleavage site that intervenes between the donor fluorophore and the acceptor, where resonance energy transfer is exhibited between the donor fluorophore and the acceptor under the appropriate conditions; (b) exciting the donor fluorophore; and (c) determining resonance energy transfer of the contacted cell relative to a control cell, where a difference in resonance energy transfer of the contacted cell as compared to the control cell is indicative of clostridial toxin activity.
摘要翻译： 本发明提供了一种通过（a）使样品与包含供体荧光团的梭菌毒素底物的细胞接触来测定梭菌毒素活性的方法; 具有与供体荧光团的发射光谱重叠的吸收光谱的受体; 以及包含介入供体荧光团和受体之间的切割位点的梭菌毒素识别序列，其中在适当条件下在供体荧光团和受体之间显示共振能量转移; （b）激发供体荧光团; 和（c）确定接触细胞相对于对照细胞的共振能量转移，其中与对照细胞相比，所接触的细胞的共振能量转移的差异指示梭菌毒素活性。
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公开(公告)号：US08535941B2
公开(公告)日：2013-09-17
申请号：US13406224
申请日：2012-02-27
CPC分类号： G01N33/5014 , G01N33/5058 , G01N33/542 , G01N33/56911 , G01N33/94
摘要： Compositions useful for detecting Clostridial toxin activity comprising a cell that comprises a membrane-associated Clostridial toxin substrate comprising a first member of a fluorescence resonance energy transfer pair; and a Clostridial toxin recognition sequence including a cleavage site; and a membrane-associated second member of the FRET pair and methods useful for determining Clostridial toxin activity using such Clostridial toxin substrates.
摘要翻译： 用于检测梭菌毒素活性的组合物，其包含细胞，所述细胞包含与所述膜相关的梭菌毒素底物，所述底物包含荧光共振能量转移对的第一成员; 和包含切割位点的梭菌毒素识别序列; 和FRET对的膜相关的第二个成员以及可用于使用这种梭菌毒素底物测定梭菌毒素活性的方法。
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57.发明申请公开(公告)号：US20120231538A1
公开(公告)日：2012-09-13
申请号：US13406224
申请日：2012-02-27
IPC分类号： C12N5/10
CPC分类号： G01N33/5014 , G01N33/5058 , G01N33/542 , G01N33/56911 , G01N33/94
摘要： Compositions useful for detecting Clostridial toxin activity comprising a cell that comprises a membrane-associated Clostridial toxin substrate comprising a first member of a fluorescence resonance energy transfer pair; and a Clostridial toxin recognition sequence including a cleavage site; and a membrane-associated second member of the FRET pair and methods useful for determining Clostridial toxin activity using such Clostridial toxin substrates.
摘要翻译： 用于检测梭菌毒素活性的组合物，其包含细胞，所述细胞包含与所述膜相关的梭菌毒素底物，所述底物包含荧光共振能量转移对的第一成员; 和包含切割位点的梭菌毒素识别序列; 和FRET对的膜相关的第二个成员以及可用于使用这种梭菌毒素底物测定梭菌毒素活性的方法。
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公开(公告)号：US08067231B2
公开(公告)日：2011-11-29
申请号：US11908451
申请日：2006-04-04
CPC分类号： G01N33/56911 , G01N2333/33
摘要： Compositions useful for detecting Clostridial toxin activity comprising a cell that contains an exogenous Clostridial toxin substrate which comprises a fluorescent member, a membrane targeting domain and a Clostridial toxin recognition sequence comprising a cleavage site, where the cleavage site intervenes between the fluorescent member and the membrane targeting domain; and methods useful for determining Clostridial toxin activity using such Clostridial toxin substrates.
摘要翻译： 用于检测梭菌毒素活性的组合物，其包含含有外源性梭菌毒素底物的细胞，其包含荧光成员，膜靶向结构域和包含切割位点的梭菌毒素识别序列，其中所述切割位点介于荧光部件和膜之间 定位域; 以及用于使用这种梭菌毒素底物测定梭菌毒素活性的方法。
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59.发明申请公开(公告)号：US20100280222A1
公开(公告)日：2010-11-04
申请号：US11620579
申请日：2007-01-05
IPC分类号： C07K14/33
摘要： Natural and modified neurotoxins and isolated neurotoxin compositions are described. The neurotoxins may include one or more structural modifications, wherein the structural modification(s) alters the biological persistence, such as the biological half-life and/or a biological activity of the modified neurotoxin relative to an identical neurotoxin without the structural modification(s). In one embodiment, methods of making the modified neurotoxin include using recombinant techniques. In another embodiment, methods of using the modified neurotoxin to treat conditions include treating various disorders, neuromuscular aliments and pain.
摘要翻译： 描述了天然和修饰的神经毒素和分离的神经毒素组合物。 神经毒素可以包括一种或多种结构修饰，其中结构修饰改变了相对于相同的神经毒素而没有结构修饰的生物学持久性，例如修饰的神经毒素的生物半衰期和/或生物活性 ）。 在一个实施方案中，制备修饰的神经毒素的方法包括使用重组技术。 在另一个实施方案中，使用经修饰的神经毒素治疗病症的方法包括治疗各种疾病，神经肌肉营养和疼痛。
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公开(公告)号：US20100081157A1
公开(公告)日：2010-04-01
申请号：US12620417
申请日：2009-11-17
IPC分类号： C12Q1/37
CPC分类号： C12Q1/37 , C07K14/001 , C07K14/435 , G01N33/56911 , G01N2333/33
摘要： The present invention provides clostridial toxin substrates useful in assaying for the protease activity of any clostridial toxin, including botulinum toxins of all serotypes as well as tetanus toxins. A clostridial toxin substrate of the invention contains a donor fluorophore; an acceptor having an absorbance spectrum overlapping the emission spectrum of the donor fluorophore; and a clostridial toxin recognition sequence that includes a cleavage site, where the cleavage site intervenes between the donor fluorophore and the acceptor and where, under the appropriate conditions, resonance energy transfer is exhibited between the donor fluorophore and the acceptor.
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