Controlled Immobilization Strategies to Probe Short Hyaluronan-Protein Interactions

Abstract

Well-controlled grafting of small hyaluronan oligosaccharides (sHA) enables novel approaches to investigate biological processes such as angiogenesis, immune reactions and cancer metastasis. We develop two strategies for covalent attachment of sHA, a fast high-density adsorption and a two-layer system that allows tuning the density and mode of immobilization. We monitored the sHA adlayer formation and subsequent macromolecular interactions by label-free quartz crystal microbalance with dissipation (QCM-D). The modified surfaces are inert to unspecific protein adsorption, and yet retain the specific binding capacity of sHA. Thus they are an ideal tool to study the interactions of hyaluronan-binding proteins and short hyaluronan molecules as demonstrated by the specific recognition of LYVE-1 and aggrecan. Both hyaladherins recognize sHA and the binding is independent to the presence of the reducing end.

Multivalent and multifunctional polysaccharide-based particles for controlled receptor recognition

“…1b ). Besides HA 49 and LAM 50 , other polysaccharides such as xylan, pullulan, dextran or galactan can also be functionalized in the same way 51 – 55 . The two building blocks were then associated by Cu(I)-catalyzed Huisgen cycloaddition as shown in Fig.…”

Supramolecular Presentation of Hyaluronan onto Model Surfaces for Studying the Behavior of Cancer Stem Cells

“…QCM-D experiments on the immobilization of alginate (a negatively charged polysaccharide) on Pep-1 SAMs showed mass deposition (Δm = 2807.5 ± 192.5 ng cm −2 ) that was slightly removed upon washing ( Figure S9 fluorescence. [16] In another study, BSA (1 mg mL −1 ) adsorption onto surfaces spin-coated with photoreactive HA (modified with 4-azidoaniline) and UV-crosslinked, monitored by QCM-D, revealed significant levels of albumin adsorption (≈250 ng cm −2 ). The next step was to gain insights on the interactions of HA surfaces with serum proteins, which is key to understand cell behavior on these surfaces.…”

“…[11,12] It has been demonstrated that signalling downstream of HA-CD44 interactions can regulate stem cell markers and highlights the role of HA in HNSCC progression. These approaches often require the chemical modification of native HA by biotinylation, [14] conjugation to dopamine, [15] thiolation, [16] azidation, [17] or functionalization of surfaces to allow the covalent attachment of HA. These approaches often require the chemical modification of native HA by biotinylation, [14] conjugation to dopamine, [15] thiolation, [16] azidation, [17] or functionalization of surfaces to allow the covalent attachment of HA.…”

The interaction of protein-coated bionanoparticles and surface receptors reevaluated: how important is the number of bonds?

“…Furthermore, the decreasing size of the lateral distance is small enough to be negligible within the timeframe of our experiments of around hundreds of seconds. The decrease of the thickness of the diffusional boundary layer corresponds to a lower adsorption rate (see eqn (1)). The decrease of this adsorption rate in turn is due to a lower availability of receptors because of their occupation by bound bionanoparticles.…”

“…d(x) is the thickness of the velocity boundary layer. v x (y), eqn (4), is the velocity of the fluid in the velocity boundary-layer region and v e (x), eqn(1) and(2), is the velocity of the fluid in the potential flow region between two velocity boundary layers. Velocity profiles at three different positions are depicted.…”