Protein adsorption and interactions with mesoporous silica are of interest for a broad range of applications including drug delivery, chemical synthesis, biosensors, and bioseparations. A major challenge in designing mesoporous silica supports for tailored protein interaction is the differentiation of protein interactions at the surface of the particle from interactions within the pore, important features when considering mesoporous silica as a protective support for active proteins. In this investigation, the location of Enhanced Green Fluorescent Proteins (EGFPs) adsorbed on tailored mesoporous silica particles is examined as a function of pore diameter using proteolytic hydrolysis to distinguish between accessible and inaccessible proteins. Pore size control is achieved by tuning the hydrothermal aging temperature (60-110 °C) during synthesis, where the synthesis results in 5-15 μm diameter spherical particles appropriate for imaging by confocal scanning laser microscopy (CSLM). In low pH environments, EGFP unfolds within pores and on the surface of particles, rendering it susceptible to proteolytic hydrolysis by the protease Pepsin A. Upon return to neutral pH, un-hydrolyzed EGFP regains its fluorescence and can be visualized within the mesoporous particles. The pore-size dependent loading and protection of EGFP (2.4 nm diameter×4.2 nm) from proteolytic attack by Pepsin A (7.3 nm×3.6 nm×5.4 nm) is demonstrated by the retention of fluorescence in 7.3 nm pores. Larger-pored materials (>9 nm) provide diminishing protection for EGFP, and the protection is greatly reduced with increasing pore size and pore size distribution breadth. Proteolytic hydrolysis is used to delineate the activity of pore-loaded versus surface-bound proteins and to establish that there is an optimal pore diameter for loading EGFP while protecting it from attack by a larger proteolytic enzyme.

Incorporation of lipid assemblies on the surface and within pores of mesoporous silica particles provides for biomimetic approaches to analyte sensing and separations using high surface area platforms. This work investigates the effect of pore confinement on the location and the diffusivity of lipid assemblies in mesoporous silica spherical particles (SBAS) as a function of nanopore diameters (nonporous, 3.0, 5.4, and 9.1 nm), which span the range of the thickness of the 1,2‐dipalmitoyl‐sn‐glycero‐3‐phosphocholine lipid bilayer (≈4 nm). Large‐diameter SBAS are imaged with sufficient spatial resolution to distinguish lipids at the exterior surface and in the center of the particles. Lipids incorporated on the silica by evaporation deposition exist as exterior lipid bilayers on all particles and lipid assemblies in the pores of 5.4 and 9.1 nm pore diameter materials. Lipid diffusivity increases with pore size and decreases in the presence of bilayer tethering functional groups. Lipid diffusivity in the core of the particles is similar to the surface diffusivity, consistent with long‐range mobility in accessible, ordered (but randomly oriented) mesopores of SBAS materials. This work presents a framework for interpreting high density loading of lipid bilayers and their function within mesoporous materials.

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Differentiating the chemical properties of the external and pore surface of sol-gel derived mesoporous materials by selective functionalization is important to advancing their application as platforms for biological catalysis, sensing and drug delivery. Prior selective functionalization approaches to concentrating functional groups at the external surface of particles have been limited to small pores (≤ 5.5 nm diameter) incapable of loading large biological molecules. This work investigates the selective exterior surface functionalization by amines of larger-pored (>7 nm diameter) mesoporous silica particles, which are synthesized by dual surfactant templating and hydrothermal aging. Previously developed selective functionalization techniques rely on choice of functionalization precursor, functionalization reaction time, or pore blocking (by leaving pore templates in as-synthesized materials). The effectiveness of these strategies are compared for larger-pored materials using the precursors (3aminopropyl)tris(methoxyethoxyethoxy)silane (APTMEES) and (3-aminopropyl)triethoxysilane (APTES). The extent of amine functionalization is determined as a function of precursor reaction time (10 or 20 minutes) in both as-synthesized and template-extracted materials by confocal laser

Selectively permeable biological membranes containing lipophilic barriers inspire the design of biomimetic carrier-mediated membranes for aqueous solute separation. The recovery of glucose, which can reversibly bind to boronic acid (BA) carriers, is examined in lipid pore-filled silica thin-film composite membranes with accessible mesopores. The successful incorporation of lipids (1,2-dipalmitoyl-sn-glycero-3-phosphocholine, DPPC) and BA carriers (4-((N-Boc-amino)methyl)phenylboronic acid, BAMP-BA) in the pores of mesoporous silica (∼10 nm pore diameter) through evaporation deposition is verified by confocal microscopy and differential scanning calorimetry. In the absence of BA carriers, lipids confined inside the pores of silica thin films (∼200 nm thick) provide a factor of 14 increase in diffusive transport resistance to glucose, relative to traditional supported lipid bilayers formed by vesicle fusion on the porous surface. The addition of lipid-immobilized BAMP-BA (59 mol % in DPPC) facilitates the transport of glucose through the membrane; glucose flux increases from 45 × 10 to 225 × 10 mol/m/s in the presence of BAMP-BA. Furthermore, the transport can be improved by environmental factors including pH gradient (to control the binding and release of glucose) and temperature (to adjust lipid bilayer fluidity). The successful development of biomimetic nanocomposite membranes demonstrated here is an important step toward the efficient dilute aqueous solute upgrading or separations, such as the processing of carbohydrates from lignocellulose hydrolysates, using engineered carrier/catalyst/support systems.