Protocol for preparing tyramine functionalized hyaluronic acid via aqueous DMTMM-mediated conjugation

Hyaluronic acid is a ubiquitous glycosaminoglycan with numerous biological and structural functions in the body. Here, we present a protocol for the practical and efficient functionalization of HA with tyramine. We describe the reaction conditions for its DMTMM-mediated coupling, the subsequent purification and drying steps, and product characterization through 1 H NMR and UV spectrophotometry. This protocol produces materials which can form hydrogels with tunable viscoelastic properties, for a range of biomedical applications, such as biofabrication and tissue engineering. For complete details on the use and execution of this protocol, please refer to Miklosic et al. 1 • Instructions for the amidation of hyaluronic acid with tyramine using DMTMM chemistry • Steps for product purification through precipitation, washing, and vacuum drying • Guidance on the characterization of HA-tyramine with NMR and UV spectroscopy Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Hyaluronic acid is a ubiquitous glycosaminoglycan with numerous biological and structural functions in the body. Here, we present a protocol for the practical and efficient functionalization of HA with tyramine. We describe the reaction conditions for its DMTMM-mediated coupling, the subsequent purification and drying steps, and product characterization through 1 H NMR and UV spectrophotometry. This protocol produces materials which can form hydrogels with tunable viscoelastic properties, for a range of biomedical applications, such as biofabrication and tissue engineering.