A quartz crystal microbalance method to quantify the size of hyaluronan and other glycosaminoglycans on surfaces

0566732 - ÚEM 2023 RIV DE eng J - Journal Article
Srimasorn, S. - Souter, L. - Green, D.E. - Djerbal, L. - Goodenough, A. - Duncan, J.A. - Roberts, A.R.E. - Zhang, X.L. - Debarre, D. - DeAngelis, P.L. - Kwok, Jessica - Richter, R.P.
A quartz crystal microbalance method to quantify the size of hyaluronan and other glycosaminoglycans on surfaces.
Scientific Reports. Roč. 12, č. 1 (2022), č. článku 10980. ISSN 2045-2322. E-ISSN 2045-2322
R&D Projects: GA MŠMT(CZ) EF15_003/0000419
Institutional support: RVO:68378041
Keywords : extracellular-matrix * chondroitin sulfate * grafted hyaluronan * bone-formation * QCM-D
OECD category: Biochemical research methods
Impact factor: 4.6, year: 2022
Method of publishing: Open access
https://www.nature.com/articles/s41598-022-14948-7

Hyaluronan (HA) is a major component of peri- and extra-cellular matrices and plays important roles in many biological processes such as cell adhesion, proliferation and migration. The abundance, size distribution and presentation of HA dictate its biological effects and are also useful indicators of pathologies and disease progression. Methods to assess the molecular mass of free-floating HA and other glycosaminoglycans (GAGs) are well established. In many biological and technological settings, however, GAGs are displayed on surfaces, and methods to obtain the size of surface-attached GAGs are lacking. Here, we present a method to size HA that is end-attached to surfaces. The method is based on the quartz crystal microbalance with dissipation monitoring (QCM-D) and exploits that the softness and thickness of films of grafted HA increase with HA size. These two quantities are sensitively reflected by the ratio of the dissipation shift (Delta D) and the negative frequency shift (-Delta f) measured by QCM-D upon the formation of HA films. Using a series of size-defined HA preparations, ranging in size from similar to 2 kDa tetrasaccharides to similar to 1 MDa polysaccharides, we establish a monotonic yet non-linear standard curve of the Delta D/-Delta f ratio as a function of HA size, which reflects the distinct conformations adopted by grafted HA chains depending on their size and surface coverage. We demonstrate that the standard curve can be used to determine the mean size of HA, as well as other GAGs, such as chondroitin sulfate and heparan sulfate, of preparations of previously unknown size in the range from 1 to 500 kDa, with a resolution of better than 10%. For polydisperse samples, our analysis shows that the process of surface-grafting preferentially selects smaller GAG chains, and thus reduces the average size of GAGs that are immobilised on surfaces comparative to the original solution sample. Our results establish a quantitative method to size HA and other GAGs grafted on surfaces, and also highlight the importance of sizing GAGs directly on surfaces. The method should be useful for the development and quality control of GAG-based surface coatings in a wide range of research areas, from molecular interaction analysis to biomaterials coatings.
Permanent Link: https://hdl.handle.net/11104/0338021