Abstract: Understanding protein-ligand interactions at a dynamic level is becoming progressively vital for a drug discovery, as it enables the precise identification of molecular inhibitors that selectively target proteins associated with diseases. Moving beyond static structural analysis, recent studies focus on the dynamics of binding interactions—particularly, how proteins and ligands come together and separate under different physiological conditions. Nuclear Magnetic Resonance (NMR) spectroscopy is a powerful tool, offering high-resolution insights into these interactions, revealing details of both weak and strong binding affinities by examining various chemical exchange regimes. This thesis focuses on NMR spectroscopy as an effective technique for exploring protein ligand interactions across both fast and slow exchange regimes, enhancing data quality through the use of techniques tailored for each type of binding. For fast-exchange systems with weak binding, methods such as chemical shift mapping, WaterLOGSY, and cross-saturation precisely characterize binding sites and yield dissociation constants (𝐾𝐷) by capturing population-averaged chemical shifts. Conversely, systems exhibiting strong affinities and slow exchange benefit from relaxation-based approaches like R₂ dispersion and ZZ-exchange spectroscopy, which facilitate in-depth analysis of association (𝑘𝑜𝑛) and dissociation constants (𝑘𝑜𝑓𝑓). This thesis demonstrates the necessity of employing exchange-specific techniques to ensure reliable quantitative data while exploring recent innovations that elevate the sensitivity and breadth of NMR spectroscopy in ligand screening and protein analysis.