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1. 发明授权

US08093044B2 Methods for identifying inhibitors of botulinum neurotoxins 有权
标题翻译：鉴定肉毒杆菌神经毒素抑制剂的方法
公开(公告)号：US08093044B2
公开(公告)日：2012-01-10
申请号：US12630336
申请日：2009-12-03
申请人： Randall Kincaid , George Oyler , Yien Che Tsai , Paul S. Fishman
发明人： Randall Kincaid , George Oyler , Yien Che Tsai , Paul S. Fishman
IPC分类号： C07K14/00 , A61K39/08 , C12N5/10
CPC分类号： C07K14/705 , C12Q1/37 , G01N33/5014 , G01N2500/04
摘要： A system and method for identifying a botulinum neurotoxin inhibitor employing a botulinum neurotoxin substrate complex having a peptide substrate, preferably SNAP-25, a reporter domain on one side of said peptide substrate and an immobilization domain on the opposite side of said peptide substrate. The botulinum neurotoxin inhibitor is identified by its ability to decrease the relative amount of cleaved complex, detected through measuring a decrease in complex bound to a solid support. The method of the present invention also utilizes novel cells that express a botulinum neurotoxin substrate complex. The methods of the present invention are adapted for cell based screening to monitor the catalytic activity of a BoNT in living cells and to identify molecules that inhibit the catalytic activity of a BoNT in living cells. Also provided are novel stable cell lines that express the botulinum toxin substrate complex and viral vectors capable of efficiently expressing an active light chain of the BoNT within mammalian cells.
摘要翻译：一种使用具有肽底物的肉毒杆菌神经毒素底物复合物鉴定肉毒杆菌神经毒素抑制剂的系统和方法，优选SNAP-25，所述肽底物一侧的报道结构域和所述肽底物相反侧的固定结构域。肉毒杆菌神经毒素抑制剂通过其降低切割复合物的相对量的能力来鉴定，通过测量与固体支持物结合的复合物的降低来检测。本发明的方法还利用表达肉毒神经毒素底物复合物的新细胞。本发明的方法适用于基于细胞的筛选以监测活细胞中BoNT的催化活性并鉴定抑制活细胞中BoNT的催化活性的分子。还提供了表达肉毒杆菌毒素底物复合物的新型稳定细胞系和能够在哺乳动物细胞内有效表达BoNT活性轻链的病毒载体。

2. 发明申请

US20060233836A1 Methods for identifying inhibitors of botulinum neurotoxins 失效
公开(公告)号：US20060233836A1
公开(公告)日：2006-10-19
申请号：US11095055
申请日：2005-03-31
申请人： Randall Kincaid , George Oyler , Yien Tsai , Paul Fishman
发明人： Randall Kincaid , George Oyler , Yien Tsai , Paul Fishman
IPC分类号： A61K39/08 , G01N33/554 , G01N33/569 , C07K14/33
CPC分类号： C07K14/705 , C12Q1/37 , G01N33/5014 , G01N2500/04
摘要： A system and method for identifying a botulinum neurotoxin inhibitor employing a botulinum neurotoxin substrate complex having a peptide substrate, preferably SNAP-25, a reporter domain on one side of said peptide substrate and an immobilization domain on the opposite side of said peptide substrate. The botulinum neurotoxin inhibitor is identified by its ability to decrease the relative amount of cleaved complex, detected through measuring a decrease in complex bound to a solid support. The method of the present invention also utilizes novel cells that express a botulinum neurotoxin substrate complex. The methods of the present invention are adapted for cell based screening to monitor the catalytic activity of a BoNT in living cells and to identify molecules that inhibit the catalytic activity of a BoNT in living cells. Also provided are novel stable cell lines that express the botulinum toxin substrate complex and viral vectors capable of efficiently expressing an active light chain of the BoNT within mammalian cells.

3. 发明申请

US20080076161A1 Method of designing synthetic nucleic acid sequences for optimal protein expression in a host cell 审中-公开
标题翻译：设计用于在宿主细胞中最佳蛋白质表达的合成核酸序列的方法
公开(公告)号：US20080076161A1
公开(公告)日：2008-03-27
申请号：US11907584
申请日：2007-10-15
申请人： Evelina Angov , Jeffrey Lyon , Randall Kincaid
发明人： Evelina Angov , Jeffrey Lyon , Randall Kincaid
IPC分类号： C12P19/34 , C12N1/20
CPC分类号： C07K14/445 , A61K39/00 , C12N15/67 , C12P21/02 , Y02A50/412
摘要： The present invention provides a method for modifying a wild type nucleic acid sequence encoding a polypeptide to enhance expression and accumulation of the polypeptide in the host cell by harmonizing synonymous codon usage frequency between the foreign DNA and the host cell DNA. This can be done by substituting codons in the foreign coding sequence with codons of similar usage frequency from the host DNA/RNA which code for the same amino acid. The present invention also provides novel synthetic nucleic acid sequences prepared by the method of the invention.
摘要翻译：本发明提供了通过调节外源DNA和宿主细胞DNA之间的同义密码子使用频率来修饰编码多肽的野生型核酸序列以增强多肽在宿主细胞中的表达和积累的方法。这可以通过用来自编码相同氨基酸的宿主DNA / RNA的具有相似使用频率的密码的外源编码序列中的密码子来代替。本发明还提供了通过本发明的方法制备的新型合成核酸序列。

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