We have developed an entirely new assay principle, the supramolecular tandem assays, in which supramolecular host molecules are employed and compete with enzymes for the substrate or the product. This new economic, convenient, and general assay principle is based on the reversible interaction of water-soluble macrocycles and fluorescent dyes. We have shown that an assortment of enzymatic reactions may be continuously monitored by measuring changes in fluorescence, which result from the competition of the enzymatic product and the dye for forming a complex with a cucurbituril or calixarene macrocycle. The new assay provides a complementary method to the use of antibodies, radioactive markers, and labeled substrates. Until now, we could assay five out of six enzyme classes (EC) with this assay principle including medically important enzymes such as methyltransferases, acetylcholine esterase, and proteases.
Key references: (a) Nat. Meth. 2007, 4, 629-632; (b) Chem. Eur. J. 2012, 18, 3444-3459; (c) Supramolecular Enzyme Assays, in: Supramolecular Systems in Biomedical Fields, 2012, ed.: H.-J. Schneider, RSC Publishing, pp. 355-396.