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A highly sensitive and class-specific fluorescence polarisation assay for sulphonamides based on dihydropteroate
Zhanhui Wang1, Xiao Liang2, Kai Wen1
1College of Veterinary Medicine, China Agricultural University, Beijing Laboratory for Food Quality and Safety, Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, Beijing 100193, People's Republic of China.
Abstract:
We describe a fluorescence polarisation assay based on the use of dihydropteroate synthase (DHPS) and a fluorescence probe for multi-sulphonamide detection. Dihydropteridine pyrophosphate (DHPPP) was synthesised and acts as the first substrate for DHPS. Under optimised conditions, the half-maximal inhibitory concentrations (IC50) of the assay were less than 100 ng mL(-1) for at least 29 sulphonamides and the time needed for the detection was less than 20 min. More importantly, the assay revealed quite uniform affinities for all of the individual sulphonamides tested, which has never before been achieved in an antibody-based assay.
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