MEASUREMENT OF MOLECULAR INTERACTIONS AT SINGLE MOLECULE LEVEL USING SUBSTRATES THAT ENHANCE FLUORESCENCE DETECTION

The invention relates to methods for characterizing the binding interactions between binding partners. The invention pertains to methods comprising contacting a first binding partner with a second binding partner that exhibits fast-off rate binding characteristics with the first binding partner to generate the binding interaction between the first binding partner and the second binding partner, wherein the first binding partner is immobilized onto a substrate that is designed to enhance the fluorescence signal of fluorescent molecules located near the substrate's surface, and wherein the second binding partner is a molecule that emits a fluorescent signal or is conjugated to molecule that emits a fluorescent signal. The interactions between the two binding partners can be analyzed based on multiple transient interactions between the two binding partners.