公开(公告)号：US07393443B2

公开(公告)日：2008-07-01

申请号：US10924235

申请日：2004-08-23

申请人： Wilfredo Colón ,  Marta Manning

发明人： Wilfredo Colón ,  Marta Manning

IPC分类号： B01D59/42 ,  G01N31/00

CPC分类号： G01N33/6803 ,  G01N2500/00

摘要： The present invention provides a fast and efficient means for identifying kinetically stable proteins. As used herein the term “kinetically stable protein” means a protein that is trapped in a specific conformation due to an unusually high unfolding barrier that results in very slow unfolding rates. The present inventors are the first to discover the existence of a correlation between kinetic stability and SDS-induced denaturation. Thus, the invention provides methods for identifying kinetically stable proteins comprising the step of testing the proteins for resistance to denaturation by SDS. In one embodiment, SDS-polyacrylamide gel electrophoresis (SDS-PAGE) is one simple method for quickly identifying and selecting kinetically stable proteins.

摘要翻译： 本发明提供用于鉴定动力学稳定蛋白质的快速和有效的方法。 如本文所用，术语“动力学稳定蛋白”是指由于非常高的展开屏障导致非常缓慢的展开速率而被捕获在特定构象中的蛋白质。 本发明人首先发现动力学稳定性与SDS诱导的变性之间存在相关性。 因此，本发明提供了用于鉴定动力学稳定蛋白质的方法，其包括测试蛋白质以抵抗SDS变性的步骤。 在一个实施方案中，SDS-聚丙烯酰胺凝胶电泳（SDS-PAGE）是快速鉴定和选择动力学稳定蛋白质的简单方法。

公开(公告)号：US07217348B2

公开(公告)日：2007-05-15

申请号：US11226072

申请日：2005-09-14

申请人： Wilfredo Colón ,  Marta Manning

发明人： Wilfredo Colón ,  Marta Manning

IPC分类号： B01D57/02

CPC分类号： G01N33/6803 ,  G01N2500/00

摘要： The present invention provides a fast and efficient means for identifying kinetically stable proteins. As used herein the term “kinetically stable protein” means a protein that is trapped in a specific conformation due to an unusually high unfolding barrier that results in very slow unfolding rates. The present inventors are the first to discover the existence of a correlation between kinetic stability and SDS-induced denaturation. Thus, the invention provides methods for identifying kinetically stable proteins comprising the step of testing the proteins for resistance to denaturation by SDS. In one embodiment, SDS-polyacrylamide gel electrophoresis (SDS-PAGE) is one simple method for quickly identifying and selecting kinetically stable proteins. In another embodiment a two-dimensional SDS-PAGE provides a high throughput method for quickly identifying kinetically stable proteins in a sample.

摘要翻译： 本发明提供用于鉴定动力学稳定蛋白质的快速和有效的方法。 如本文所用，术语“动力学稳定蛋白”是指由于非常高的展开屏障导致非常缓慢的展开速率而被捕获在特定构象中的蛋白质。 本发明人首先发现动力学稳定性与SDS诱导的变性之间存在相关性。 因此，本发明提供了用于鉴定动力学稳定蛋白质的方法，其包括测试蛋白质以抵抗SDS变性的步骤。 在一个实施方案中，SDS-聚丙烯酰胺凝胶电泳（SDS-PAGE）是快速鉴定和选择动力学稳定蛋白质的简单方法。 在另一个实施方案中，二维SDS-PAGE提供用于快速鉴定样品中的动力学稳定蛋白质的高通量方法。