公开(公告)号：US20140287433A1

公开(公告)日：2014-09-25

申请号：US14233740

申请日：2012-07-18

申请人： Oliver Weingart ,  Marc Avondet ,  Martin Loessner ,  Andreas Rummel ,  Frank Gessler

发明人： Oliver Weingart ,  Marc Avondet ,  Martin Loessner ,  Andreas Rummel ,  Frank Gessler

IPC分类号： G01N33/573

CPC分类号： G01N33/573 ,  G01N33/5432 ,  G01N2333/33 ,  G01N2333/952

摘要： This invention relates to a method of determining presence, amount and/or activity of a clostridial neurotoxin in a sample, the method comprising or consisting of the following steps: (a) bringing said sample into contact with a liposome, said liposome comprising (aa) at least one receptor on its outer surface, said receptor being capable of binding said neurotoxin and comprising or consisting of (i) a glycolipid and (ii) a peptide or protein; and (ab) a substrate in its interior, said substrate (i) being cleavable by the peptidase comprised in said neurotoxin and (ii) generating a detectable signal upon cleavage, said detectable signal preferably being generated by (1) the donor of a FRET pair, said donor exhibiting increased fluorescence upon cleavage by said peptidase, (2) a luminescent compound formed upon said cleavage, or (3) an enzyme formed upon said cleavage; and (b) determining whether an increase in signal occurs as compared to the absence of said sample, wherein such increase is indicative of the presence of said neurotoxin and/or the degree of such increase is indicative of the amount and/or activity of said neurotoxin in said sample.

摘要翻译： 本发明涉及一种测定样品中梭菌神经毒素的存在，量和/或活性的方法，所述方法包括或由以下步骤组成：（a）使所述样品与脂质体接触，所述脂质体包含（aa ）在其外表面上的至少一个受体，所述受体能够结合所述神经毒素并且包含（i）糖脂和（ii）肽或蛋白质或由其组成; 和（ab）其内部的底物，所述底物（i）可由包含在所述神经毒素中的肽酶切割，和（ii）在切割时产生可检测信号，所述可检测信号优选通过（1）FRET的供体产生 所述供体在所述肽酶切割时表现出增加的荧光，（2）在所述切割时形成的发光化合物，或（3）在所述切割时形成的酶; 和（b）确定与不存在所述样品相比是否发生信号增加，其中这种增加指示所述神经毒素的存在和/或这种增加的程度指示所述神经毒素的量和/或活性 所述样品中的神经毒素。