NMR spectroscopy is a powerful technique for probing protein structure, stability, function and folding. Any expansion of NMR to other Γelds, such as engineering, could only be a beneΓt. Since protein thermal stability studies can provide useful information for protein processing in the biopharmaceutical industry, the applicability of one-dimensional proton (1D 1H) NMR for thermal denaturation midpoint and longitudinal relaxation times (T m ) for thermal stability data were investigated for the globular protein lysozyme. The considerations for protein (T 1 ) NMR spectral analysis through temperature changes are elaborated upon. These considerations may also be of great value for protein, peptide, carbohydrate and oligonucleotide spectral analysis through changes other than temperature, such as pressure, pH, concentration and solute interactions. Since the experimental considerations elaborated upon were used for both and analysis, they should also apply to the determination of other T 1 T m NMR-derived data. The denatured and native H15 C2H hen lysozyme protons were used to estimate to within T m 2.3% . The native H15 C2H 1H is greater than the denatured 1H at all temperatures ascertained in the T 1 T 1 presence or absence of polyethylene glycol 1000 (PEG 1000). The di β erence in the native and denatured H15 C2H values indicates that the surface-located histidine residue experiences a di β ering local native conformation than T 1 the denatured local averaged confomation state. The H15 C2H denatured 1H