Ultrasensitive Detection of Aminopeptidase N Activity in Urine and Cells with a Ratiometric Fluorescence Probe.

An ultrasensitive ratiometric fluorescent probe (CVN) has been designed and synthesized by incorporating alanine into the cresyl violet fluorophore. The probe shows ratiometric fluorescence response toward aminopeptidase N (APN) through the increase of fluorescent intensity ratio of 626/575 nm. The sensitivity of the probe is ultrahigh with a detection limit of 33 pg/mL, which can quantify the contents of APN in 500-fold diluted human urine samples. Furthermore, by using ratiometric fluorescence imaging, the probe reveals significantly higher contents of APN in HepG2 cells than those in LO2 cells, which has been further used to distinguish these two types of cells in mixed cocultures. The probe could be of great importance for the APN-related disease diagnosis and pathophysiology elucidation.