Targeted Analysis of the Size Distribution of Heavy Chain-Modified Hyaluronan with Solid-State Nanopores

TY - JOUR

T1 - Targeted Analysis of the Size Distribution of Heavy Chain-Modified Hyaluronan with Solid-State Nanopores

AU - Erxleben, Dorothea A

AU - Dodd, Rebecca J

AU - Day, Anthony J

AU - Green, Dixy E

AU - DeAngelis, Paul L

AU - Poddar, Suruchi

AU - Enghild, Jan J

AU - Huebner, Janet L

AU - Kraus, Virginia B

AU - Watkins, Amanda R

AU - Reesink, Heidi L

AU - Rahbar, Elaheh

AU - Hall, Adam R

PY - 2024/1/30

Y1 - 2024/1/30

N2 - 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 (HC•HA) complexes. While limited assessments of HC•HA have been performed previously, determining the size distribution of its HA component remains a challenge. Here, we describe a selective method for extracting HC•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 HC•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 HC•HA and total HA in an equine model of synovitis.

AB - 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 (HC•HA) complexes. While limited assessments of HC•HA have been performed previously, determining the size distribution of its HA component remains a challenge. Here, we describe a selective method for extracting HC•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 HC•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 HC•HA and total HA in an equine model of synovitis.

KW - Alpha-Globulins/metabolism

KW - Animals

KW - Horses

KW - Humans

KW - Hyaluronic Acid/chemistry

KW - Inflammation

KW - Nanopores

KW - Synovial Fluid

U2 - 10.1021/acs.analchem.3c04387

DO - 10.1021/acs.analchem.3c04387

M3 - Journal article

C2 - 38215004

SN - 0003-2700

VL - 96

SP - 1606

EP - 1613

JO - Analytical Chemistry

JF - Analytical Chemistry

IS - 4

ER -