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This lecture covers the classifications and configurations of molecular assays, including homogeneous and heterogeneous assays such as direct, sandwich, competition, and inhibition assays. It discusses the main issues encountered in surface sensing, such as reagent availability, mass-transport phenomena, interference in matrices, ligand design, stability, and regeneration. The lecture also delves into molecular quantification, explaining the upper and lower limits of measurable concentrations and the impact of excess reagent conditions. Additionally, it explores the challenges related to target molecule availability, interference from non-specific molecules, and the kinetics of binding and dissociation on surfaces.