A 'turn-off' fluorescent substrate for horseradish peroxidase detects anti-chlamydia in mouse serum by enhancing accuracy of ELISA kits by 10 fold

摘要

The variability in detection of antigens in human tissue samples via ELISAs is a central problem in medicine and limits diagnosis of otherwise detectable disorders. In this report we present a fluorescent "turn-off' ultrasensitive HRP substrate termed, cLiCs (cyanine Liposome Complexes), which increase the accuracy of protein detection using commercial ELISA kits by 20 fold because they reduce the variation due to non-specific background signal. cLiCs have been designed to decrease the diffusion distance between reactive oxygen species (ROS) enhancers and the ROS reactive cyanine dyes to nanometer scale by localizing them in a lipid bilayer, and this allows for increase in sensitivity and detection of as few as 100,000 molecules of soluble HRP in a 96 well plate. We show here that cLiCs detect anti-chlamydia antibodies in mouse serum and commercial TMB substrates could not detect these antibodies. The high accuracy of cLiCs combined with their compatibility with commercial ELISA kits gives them the ability to impact numerous areas of biology and medicine.