Selective external surface functionalization of large-pore silica materials capable of protein loading

Differentiating the chemical properties of the external and pore surfaces 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 (3-aminopropyl)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 min) in both as-synthesized and template-extracted materials by confocal laser scanning microscopy of the ∼10 μm diameter particles tagged with fluorescein isothiocyanate. Reaction time, regardless of pore template presence, is demonstrated to be the controlling variable for achieving selective exterior functionalization in these larger pored mesoporous materials. Under the conditions used, 10 min of functionalization with APTMEES localizes amine groups at the exterior of the particles, while 20 min functionalizes both the exterior and the interior pore surfaces. Protein accessibility within pores, before and after selective and full functionalization, is visually confirmed by confocal fluorescence imaging of Rhodamine B tagged lysozyme loaded particles.