申请人:Design-Zyme LLC
公开号:US11021730B2
公开(公告)日:2021-06-01
The present invention is broadly concerned with new in vitro glycosylation methods that provide rational approaches for producing glycosylated proteins, and the use of glycosylated proteins. In more detail, the present invention comprises methods of glycosylating a starting protein having an amino sidechain with a nucleophilic moiety, comprising the step of reacting the protein with a carbohydrate having an oxazoline moiety on the reducing end thereof, to covalently bond the amino sidechain of the starting protein with the oxazoline moiety, wherein the glycosylated protein substantially retains the structure and function of the starting protein. Target proteins include oxidase, oxidoreductase and dehydrogenase enzymes. The glycosylated proteins advantageously have molecular weights of at least about 7500 Daltons. In a further embodiment, the present invention concerns the use of glycosylated proteins, fabricated by the methods disclosed herein, in the assembly of amperometric biosensors.
本发明广泛涉及新的体外糖基化方法,该方法提供了生产糖基化蛋白质的合理方法,以及糖基化蛋白质的用途。更详细地说,本发明包括对具有亲核分子氨基侧链的起始蛋白质进行糖基化的方法,包括使蛋白质与还原端具有噁唑啉分子的碳水化合物反应,使起始蛋白质的氨基侧链与噁唑啉分子共价键合,其中糖基化的蛋白质基本上保留了起始蛋白质的结构和功能。目标蛋白质包括氧化酶、氧化还原酶和脱氢酶。糖基化蛋白质的分子量至少约为 7500 道尔顿。在另一个实施方案中,本发明涉及糖基化蛋白质在安培生物传感器组装中的使用,糖基化蛋白质是通过本文公开的方法制造的。