Mechanosensitivity in biology, e.g., cells responding to material stiffness, is important for the design of synthetic biomaterials. It is caused by protein receptors able to undergo conformational changes depending on mechanical stress during adhesion processes. Here the elastic modulus dependence of adhesive interactions is systematically quantified using ligand-receptor model systems that are generally not thought to be mechanosensitive: biotinavidin, mannose-concanavalin A, and electrostatic interactions between carboxylic acids and polycationic surfaces. Interactions are measured by microgel sensors of different stiffness adhering to surfaces presenting a corresponding binding partner. Adhesion is generally decreased for softer microgels due to reduced density of binding partners. Density-normalized data show that low-affinity carbohydrate ligands exhibit reduced binding in softer networks, probably due to increased network conformational entropy. However, in case of stronger interactions with large interaction range (electrostatic) and large lifetime (biotin-avidin) density normalized adhesion isincreased. This suggests compensation of entropic repulsion for softer networks probably due to their increased mechanical deformation upon microgel adhesion and enhanced cooperative binding. In essence, experiments indicate that soft interacting polymer materials exhibit entropic repulsion, which can be overcome by strongly interacting species in the network harnessing network flexibility in order to increase adhesion.
We present a synthetic approach toward soft, glycooligomer-functionalized microgel particles mimicking carbohydrate presenting cell surfaces and analyze their specific binding to a model lectin (Concanavalin A, ConA). Focusing on multivalent presentation, a series of sequence-controlled glycooligomers with varying spacing and number of mannose units was synthesized and analyzed for the resulting glycooligomer-ConA affinity. Both direct binding and inhibition studies show a higher affinity with increasing the number of sugar moieties, but they level off for higher valent systems, indicating steric hindrance. Furthermore, the results suggest that increasing the scaffold length tends to decrease binding due to entropic repulsion, which could be compensated by larger scaffolds able to address multiple ConA binding sites. These findings were consistent in all assays (adhesion, fluorescence, and ITC) regardless of binding partner immobilization, demonstrating that flexible ligands exert similar binding modes in solution and when attached to polymer networks, which is relevant for designing glyco-functionalized materials.
A synthesis toward sequence-controlled multiblock glycopolymers, presenting a mannopyranoside (Man) glyco(oligoamide) block followed by a poly(ethylene glycol) (PEG) (M̅ n of 6 kDa) block, is shown. Therefore, monodisperse and sequence-defined glyco(oligoamide) macromonomers derived from solid phase synthesis (SPS) are polymerized with dithiol-functionalized PEG via thiol–ene coupling (TEC) in a step-growth fashion. For the polymerization, a novel building block introducing a norbornene moiety is developed which is used for end-functionalization of the glyco(oligoamide) macromonomers. As a highly reactive alkene moiety in photoinduced TEC, this gives access to X̅ n of up to 45. A total of 12 glyco(oligoamide)–PEG multiblock copolymers with maximum M̅ n of 200 kDa are obtained and subjected to a series of purification steps decreasing overall dispersity. In different binding studies toward model lectin Concanavalin A, despite their high number of Man ligands, we see rather weak binding of glycopolymers that we attribute to the introduction of higher molecular weight PEG blocks.
This study aims at quantifying the steric shielding effect of multivalent glycoconjugates targeting pathogens by blocking their carbohydrate binding sites. Specifically, PEGylated and non-PEGylated glycoconjugates are studied as inhibitors of lectins and bacterial adhesins evaluating the steric repulsion effect of the nonbinding PEG chains. We use the soft colloidal probe (SCP) adhesion assay to monitor the change in the adhesion energy of mannose (Man)decorated hydrogel particles on a layer of concanavalin A (ConA) in the presence of sequence-defined multivalent glycoconjugate inhibitors over time. The results show that PEGylated glycoconjugates achieve a stronger adhesion inhibition when compared to non-PEGylated glycoconjugates although the dissociation constants (K D ) of the PEGgylated compounds to ConA were larger. These results appear in line with Escherichia coli adhesion inhibition assays showing a small increase of bacteria detachment by PEGgylated glycoconjugates compared to non-PEGylated compounds. This suggests that an increase of sterical shielding via PEGylation may help reduce the invasiveness of pathogens even after they have adhered. Adhesion studies based on electrostatic interactions using amine-linked PEG of varying molecular weight confirm that such sterical shielding effect is not limited to carbohydrate-mediated adhesion.
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