Cross-linking reactions in Langmuir monolayers of specially designed aminolipids - a toolbox for the customized production of amphiphilic nanosheets.

Synthetic amino lipids, already known as highly efficient gene therapy tool, are used in a novel way to create cross-linked stable one-molecule-thin films envisioned for future (bio)-materials applications. The films are prepared as Langmuir monolayers at the air/water interface and cross-linked 'in situ' via dynamic imine chemistry. The cross-linking process and the film characteristics are monitored by various surface-sensitive techniques such as grazing incidence X-ray diffraction, X-ray reflectivity, and infrared reflection-absorption spectroscopy. After transfer onto carbon grids, the cross-linked films are investigated by transmission and scanning electron microscopy. The obtained micrographs display mechanically self-supported nanosheets with area dimensions over several micrometers and, thus, an undeniable visual proof of successful cross-linking. The cross-linking process at the air/water interface allows to obtain Janus-faced sheets with a hydrophobic side characterized by aliphatic alkyl chains and a hydrophilic side characterized by nucleophilic groups like amines, hydroxyl groups and imine.

Membrane Adhesion via Glycolipids Occurs for Abundant Saccharide Chemistries.

Membrane-bound oligosaccharides with specific chemistries are known to promote tight adhesion between adjacent membranes via the formation of weak saccharide bonds. However, in the literature, one can find scattered evidence that other, more abundant saccharide chemistries exhibit similar behavior. Here, the influence of various glycolipids on the interaction between adjacent membranes is systematically investigated with the help of small- and wide-angle x-ray scattering and complementary neutron diffraction experiments. Added electrostatic repulsion between the membrane surfaces is used to identify the formation of saccharide bonds and to challenge their stability against tensile stress. Some of the saccharide headgroup types investigated are able to bind adjacent membranes together, but this ability has no significant influence on the membrane bending rigidity. Our results indicate that glycolipid-mediated membrane adhesion is a highly abundant phenomenon and therefore potentially of great biological relevance.

Headgroup-Ordered Monolayers of Uncharged Glycolipids Exhibit Selective Interactions with Ions.

Selective interactions of ions with charge-neutral saccharides can have far-reaching consequences in biological and wet-technological contexts but have so far been observed only indirectly. Here, we directly quantify by total-reflection X-ray fluorescence the preferential accumulation of ions near uncharged saccharide surfaces in the form of glycolipid Langmuir monolayers at air/water interfaces exhibiting different levels of structural ordering. Selective interactions with ions from the aqueous subphase are observed for monolayers featuring crystalline ordering of the saccharide headgroups, as determined by grazing-incidence X-ray diffraction. The attracted ion species depend on the structural motifs displayed by the ordered saccharide layer. Our results may constitute a basis to understand the salt-specific swelling of wood materials and various phenomena in membrane biophysics.

End Point Versus Backbone Specificity Governs Characteristics of Antibody Binding to Poly(ethylene glycol) Brushes.

End-grafted poly(ethylene glycol) (PEG) brushes are widely used in order to suppress undesired protein adsorption to surfaces exposed to blood or other biological fluids. The specific adsorption of antibodies (Abs) to PEG brushes associated with PEG's antigenicity is drawing increasing attention because it can affect clinical applications. Here, the adsorption to PEG brushes of two Ab types, specifically binding the polymer backbone and the polymer endpoints, is structurally characterized by neutron reflectometry. The measurements yield volume fraction profiles of PEG and of the adsorbed Abs with sub-nanometer resolution perpendicular to the surface. For all brush parameters in terms of grafting density and polymerization degree, the Ab profiles clearly differ between backbone binders and endpoint binders. The adsorbed Ab amount per unit area is substantial for both Ab types and for all brush parameters investigated, even for dense brushes, which impose a considerable osmotic barrier to Ab insertion. The results therefore indicate that variation of brush parameters alone is insufficient to prevent undesired Ab adsorption. Instead, our work motivates further efforts in the search for nonantigenic brush chemistry.

Conformation of Single and Interacting Lipopolysaccharide Surfaces Bearing O-Side Chains.

The outer surfaces of Gram-negative bacteria are composed of lipopolysaccharide (LPS) molecules exposing oligo- and polysaccharides to the aqueous environment. This unique, structurally complex biological interface is of great scientific interest as it mediates the interaction of bacteria with antimicrobial agents as well as with neighboring bacteria in colonies and biofilms. Structural studies on LPS surfaces, however, have so far dealt almost exclusively with rough mutant LPS of reduced molecular complexity and limited biological relevance. Here, by using neutron reflectometry, we structurally characterize planar monolayers of wild-type LPS from Escherichia coli O55:B5 featuring strain-specific O-side chains in the presence and absence of divalent cations and under controlled interaction conditions. The model used for the reflectivity analysis is self-consistent and based on the volume fraction profiles of all chemical components. The saccharide profiles are found to be bimodal, with dense inner oligosaccharides and more dilute, extended O-side chains. For interacting LPS monolayers, we establish the pressure-distance curve and determine the distance-dependent saccharide conformation.

Neutron Reflectometry Elucidates Protein Adsorption from Human Blood Serum onto PEG Brushes.

Poly(ethylene glycol) (PEG) brushes are reputed for their ability to prevent undesired protein adsorption to material surfaces exposed to biological fluids. Here, protein adsorption out of human blood serum onto PEG brushes anchored to solid-supported lipid monolayers was characterized by neutron reflectometry, yielding volume fraction profiles of lipid headgroups, PEG, and adsorbed proteins at subnanometer resolution. For both PEGylated and non-PEGylated lipid surfaces, serum proteins adsorb as a thin layer of approximately 10 Å, overlapping with the headgroup region. This layer corresponds to primary adsorption at the grafting surface and resists rinsing. A second diffuse protein layer overlaps with the periphery of the PEG brush and is attributed to ternary adsorption due to protein-PEG attraction. This second layer disappears upon rinsing, thus providing a first observation of the structural effect of rinsing on protein adsorption to PEG brushes.