RESUMEN
The glycosaminoglycan hyaluronan (HA) plays important roles in diverse physiological functions where the distribution of its molecular weight (MW) can influence its behavior and is known to change in response to disease conditions. During inflammation, HA undergoes a covalent modification in which heavy chain subunits of the inter-alpha-inhibitor family of proteins are transferred to its structure, forming heavy chain-HA (HCâ¢HA) complexes. While limited assessments of HCâ¢HA have been performed previously, determining the size distribution of its HA component remains a challenge. Here, we describe a selective method for extracting HCâ¢HA from mixtures that yields material amenable to MW analysis with a solid-state nanopore sensor. After demonstrating the approach in vitro, we validate extraction of HCâ¢HA from osteoarthritic human synovial fluid as a model complex biological matrix. Finally, we apply our technique to pathophysiology by measuring the size distributions of HCâ¢HA and total HA in an equine model of synovitis.
