Advances in NMR Spectroscopy for biological systems: Principles, techniques, and their growing scope.

Nuclear Magnetic Resonance (NMR) spectroscopy is a crucial tool in structural biology, uniquely capable of revealing protein structure, dynamics, and interactions at atomic resolution in environments that closely resemble native conditions. The combination of key methodological breakthroughs-including strategic isotopic labeling, stronger magnetic fields, cryogenic probes, and advanced pulse sequences-has established NMR as the definitive method for gaining atomic-level insights into complex biomolecules, especially pathogenic proteins involved in disease. These advances enable various NMR techniques, from high-resolution solution and solid-state NMR (ssNMR) for insoluble assemblies to in-cell NMR. Beyond structural analysis, NMR provides robust quantitative performance, high reproducibility, and rich structural information, making it a valuable platform for biomolecular analysis and metabolomics. This review aims to provide a comprehensive overview of these critical roles, with a particular emphasis on the transformative influence of integrating Artificial Intelligence (AI) into NMR techniques to accelerate metabolomics-based biomarker discovery for various diseases and conditions.