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. 2022 Jun 29;12:10980. doi: 10.1038/s41598-022-14948-7

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Standard curve for sizing HA, and extension to other GAG types. Double logarithmic plot of ΔD/− Δf (i = 3; frequency shifts are normalised by i) at low GAG surface density (− Δf = 2.5 Hz) as a function of the mean number of disaccharides per GAG-b chain, n_ds (mean ± 5%, except for oligosaccharides which were taken to be pure in size, see Table 1). Data for all size-defined HA-b samples (blue spheres) represent mean ± standard error of the mean from between two and four independent experiments (see Fig. S4 for details). The data for HA up to n_ds = 1300 are faithfully reproduced and interpolated by Eq. (1) (red line), representing the standard curve for GAG sizing. Data for size-defined preparations of other GAG types (half-filled circles; see Fig. S6 for original data) are also seen to map onto the standard curve: four heparan sulfate (HS-b; violet) oligosaccharides, two heparosan (Hep-b; orange) and one chondroitin (C0-b; dark yellow) polysaccharide. Symbols and error bars for non-HA GAGs represent individual experiments with the mean ± standard deviation time-averaged over − Δf values ranging between 2 and 3 Hz.