Oligomerization Analysis of Gal-3 Constructs

To investigate the oligomerization of different Gal-3 constructs, a Superdex 200 Increase 10/300 GL column (Cytiva, Dreieich, Germany) was used. The maximum pressure limit of the column was determined by Equations (1)–(3).
$$\mathrm{pmax}=\Delta \mathrm{p}+\Delta \mathrm{pafter}+\Delta \mathrm{pbefore}$$
$$\Delta \mathrm{p}=\Delta \mathrm{pcolumn}-\Delta \mathrm{pbefore}$$
$$\Delta \mathrm{pafter}=\Delta \mathrm{ptotal}-\Delta \mathrm{pbefore(2)}$$
Linear regression was used to calculate the molecular weights of galectin samples. For this, the K_AV (Equation (4)) of Blue Dextran and protein standards (High Molecular Weight and Low Molecular Weight; Cytiva, Germany) were plotted over the logarithmic molecular weights of the standard proteins (Figure S13). The elution volume of the samples was determined by applying 200 µL galectin sample (sterile filtered, 0.2 µM) to the column via loop injection at a constant flow rate of 0.75 mL x min⁻¹ with PBS pH 7.5 as running buffer. The elution was monitored at 280 nm.
$${\mathrm{K}}_{\mathrm{AV}}\left[-\right]=\frac{{\mathrm{V}}_{\mathrm{e}}-{\mathrm{V}}_{\mathrm{o}}}{{\mathrm{V}}_{\mathrm{c}}-{\mathrm{V}}_0}$$