SPR and ITC determination of the kinetics and the thermodynamics of bivalent versus monovalent sugar ligand–lectin interactions

Abstract

A kinetic study of the interaction of bivalent and monovalent sugar ligands with a lectin was undertaken with the aid of surface plasmon resonance (SPR) method. The study involved a series of bivalent alpha-D-mannopyranoside containing sugar ligands, with systematic variation in the distance between the sugar ligands. The detailed kinetic studies showed that bivalent ligands underwent a faster association (k(on)) and a slower dissociation (k(off)) of the ligand-lectin complexes, in comparison to the monovalent ligand-lectin complexes. The kinetic constants were complemented further by assessing the thermodynamic parameters with the aid of isothermal titration calorimetry (ITC). The initiation of cross-linking of ligand-lectin interactions emerge from the early stages of the complexation. The dynamic light scattering (DLS) and the transmission electron microscopy (TEM) techniques allowed judging the sizes and morphologies of the complex in the solution and solid states, respectively.

Synthesis and Biophysical Study of Disassembling Nanohybrid Bioconjugates with a Cubic Octasilsesquioxane Core

“…Figure 2B shows the titrations obtained for the complex formation of ligands 6c , 6d , 3c and 3d with Con A. The K D values for the monovalent compounds 6c and 6d ( Table 2 , entries 2,3) were similar to those described in the literature43–44 for methyl α ‐ D ‐mannopyranoside using this technique (Table 2, entry 1). The stoichiometry ( n ) determined for the complex of the monovalent ligands with Con A was 1:1 ( n = 1; i.e., each monovalent ligand is able to interact exclusively with one unit of Con A), as expected (Table 2, entries 2,3).…”

Exploring Structure–Property Relationships of GAGs to Tailor ECM-Mimicking Hydrogels

“…The calculated release profiles confirmed the release kinetics for the hydrogels formed out of the different heparin derivatives with a maximum of 7% PDGF-BB released for 6ON-DSH hydrogel. The dissociation constants for the PDGF-BB-heparin complexes derived from the fit of the data were in the same order of magnitude as the K d values obtained from BLI (Figure 4C) which is in agreement within the variability range of K d values when utilizing different determination methods [42,43]. Moreover, following the rational design concept for the hydrogels as morphogen delivery systems [11,13], the adjustment of the input parameters in the reaction-diffusion model such as the growth factor loading amount, the degree of sulfation, the ratio of the hydrogel volume to the release medium volume, provides the basis for the prediction of the release kinetics and morphogen gradient formation in in vitro and in vivo scenarios.…”

Analysis of Cooperativity by Isothermal Titration Calorimetry

“…Global analysis offers the possibility of combining ITC with data from other techniques such as surface plasmon resonance [50], NMR [51], circular dichroism [52], spectrofluorometry [53] and dynamic light scattering [50]. …”