Enzyme immobilization plays a crucial role in enhancing the stability and recyclability of enzymes for industrial applications. However, traditional methods for quantifying enzyme loading within porous carriers are limited by time-consuming workflows, cumulative errors, and the inability to probe enzymes adsorbed inside the pores. In this study, we introduce Time-Domain Nuclear Magnetic Resonance (TD-NMR) relaxometry as a novel, non-invasive technique for directly quantifying enzyme adsorption within porous carriers. Focusing on epoxy methyl acrylate carriers, commonly used in biocatalysis, we correlate changes in T₂ relaxation times with enzyme concentration, leading to the development of an NMR-based pore-filling ratio that quantifies enzyme loading. Validation experiments demonstrate that TD-NMR-derived adsorption curves align closely with traditional photometric measurements, offering a reliable and reproducible alternative for enzyme quantification. The accessibility of tabletop TD-NMR spectrometers makes this technique a practical and cost-effective tool for optimizing biocatalytic processes. Furthermore, the method holds promise for real-time monitoring of adsorption dynamics and could be adapted for a wider range of carrier materials and enzymes.

Stimuli-responsive gels demonstrate macroscopic changes upon exposure to external stimuli, offering potential for the development of adaptive chemical reactors. Early investigations into hydrogels established that crosslinked polymer networks experience reversible volume phase transitions, with temperature, pH, and solvent composition governing swelling and shrinking dynamics. Although hydrogels behavior in aqueous environments has been extensively characterized, lyogels that incorporate organic solvents remain comparatively underexplored, despite their potential for enhanced chemical compatibility and functional versatility. Here, we investigate how solvent polarity and crosslinking density govern the swelling behavior, pore formation, and molecular-scale dynamics of poly(N-isopropylacrylamide)-based lyogels. Using a combination of swelling measurement, scanning electron microscopy, and multiscale NMR relaxometry and diffusometry, we demonstrate that solvent polarity fundamentally alters lyogel structure and dynamics. Lyogels swollen in a high-polarity solvent exhibits macroporous networks and slower solvent exchange rates, whereas a low-polarity solvent induces shrinkage, denser microstructures, faster solvent exchange rates, and stronger surface interactions. These results establish a mechanistic framework linking thermodynamic affinity, solvent dynamics, and microstructural confinement to macroscopic gel responsiveness. This framework provides guidance for tailoring lyogels in dynamic environments, with potential applications in adaptable and tunable chemical reactors.