Polycation induced benzoperylene probe excimer formation and the ratiometric detection of heparin and heparinase.
A benzoperylene probe excimer emission in an aqueous buffer solution is observed for the first time, and a novel ratiometric fluorescence method based on the probe excimer emission for the sensitive detection of heparin and heparinase is demonstrated. A negatively charged benzoperylene derivative, 6 (benzo[ghi]perylene 1,2 dicarboxylic imide yl)hexanoic acid (BPDI) was employed. A polycation, poly(diallyldimethylammonium) chloride (poly DDA) could induce aggregation of BPDI through noncovalent interactions. A decrease of BPDI monomer emission and a simultaneous increase of BPDI excimer emission were observed. Upon the addition of heparin, the strong binding between heparin and poly DDA caused release of BPDI monomer molecules, and an excimer monomer emission signal transition was detected. However, after the enzymatic hydrolysis of heparin by heparinase, heparin was hydrolyzed into small fragments, which weakened the competitive binding of heparin to poly DDA. Poly DDA induced aggregation of BPDI, and a monomer excimer emission signal transition was detected. Our assay is simple, rapid, inexpensive, sensitive and selective, which could facilitate the heparin and heparinase related biochemical and biomedical research.
Biosensors bioelectronics