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This work addresses the influence that the nanometric features of biologically relevant surfaces have on the conformation and properties of adsorbed laminin. It was observed that the adsorption kinetics and the nanomorphology of laminin were affected by the change in local curvature of chemically homogeneous nanostructured surfaces. The nanostructured surfaces were prepared by exploiting the self-assembly process of carboxylated polystyrene NPs, with diameters of 45, 109, and 209 nm, onto a polyelectrolyte multilayer formed by alternate deposition of poly(acrylic acid) and poly(allylamine hydrochloride) on gold. The anchored polymeric NPs were finally coated with a homogeneous layer of poly(allylamine hydrochloride), providing three surfaces with different nanometric local curvature. Atomic force microscopy was employed to characterize the relevant morphological parameters of the nanostructured surfaces. Quartz crystal microbalance with dissipation monitoring was employed to determine the adsorbed mass of laminin as well as its adsorption rate as a function of the local surface curvature. A model is proposed to explain the higher and faster laminin adsorption on surfaces with lower local curvature, where a switching of laminin anchoring orientation from a side-on to an end-on geometry can be predicted by a simple curvature-dependent parameter, γ, connecting the average nanostructure height h and the macromolecule radius of gyration R(g). The results provide a framework to understand the dependence of biomolecule orientation on local nanostructure.

Broadly neutralizing anti-HIV-1 monoclonal antibody 2G12 exclusively targets a conserved cluster of high-mannose oligosaccharides present on outer viral envelope glycoprotein gp120. This characteristic makes the otherwise immunogenically "silent" glycan shield of gp120 a tempting target for drug and vaccine design. However, immune responses against carbohydrate-based mimics of gp120 have failed to provide immunization against HIV-1 infection, highlighting the need to understand the molecular events that determine immunogenicity better. In this work, the unbinding kinetics of the gp120-2G12 (k(0) = 0.002 ± 0.09 s(-1), x(++) = 1.5 ± 1.2 nm), Man(4)-2G12 (k(0) = 0.35 ± 0.32 s(-1), x(++) = 0.6 ± 0.2 nm), and Man(5)-2G12 interactions were measured by single-molecule force spectroscopy. To our knowledge, this is the first single-molecule study aimed at dissecting the carbohydrate-antibody recognition of the gp120-2G12 interaction. We were able to confirm crystallographic models that show both the binding of the linear Man(4) arm to 2G12 and also the multivalent gp120 glycan binding to 2G12. These results demonstrate that single-molecule force spectroscopy can be successfully used to dissect the molecular mechanisms underlying immunity.

The occurrence of air-trapping inside poly-eta-caprolactone nanopits was investigated by measuring the contact angles of water droplets on a set of defined nanotopographies. It is shown that the advancing angles follow the Cassie-Baxter theory, thus revealing the presence of air bubbles inside the biodegradable nanopatterns. The importance of these observations for the definition of hydrophilicity/hydrophobicity and in the context of in vitro cell behavior is discussed.