Directed evolution of artificial repeat proteins as habit modifiers for the morphosynthesis of (111)-terminated gold nanocrystals.

Natural biocomposites are shaped by proteins that have evolved to interact with inorganic materials. Protein directed evolution methods which mimic Darwinian evolution have proven highly successful to generate improved enzymes or therapeutic antibodies but have rarely been used to evolve protein-material interactions. Indeed, most reported studies have focused on short peptides and a wide range of oligopeptides with chemical binding affinity for inorganic materials have been uncovered by phage display methods. However, their small size and flexible unfolded structure prevent them from dictating the shape and crystallinity of the growing material. In the present work, a specific set of artificial repeat proteins (αRep), which exhibit highly stable 3D folding with a well-defined hypervariable interacting surface, is selected by directed evolution of a very efficient home-built protein library for their high and selective affinity for the Au(111) surface. The proteins are built from the extendable concatenation of self-compatible repeated motifs idealized from natural HEAT proteins. The high-yield synthesis of Au(111)-faceted nanostructures mediated by these αRep proteins demonstrates their chemical affinity and structural selectivity that endow them with high crystal habit modification performances. Importantly, we further exploit the protein shell spontaneously assembled on the nanocrystal facets to drive protein-mediated colloidal self-assembly and on-surface enzymatic catalysis. Our method constitutes a generic tool for producing nanocrystals with determined faceting, superior biocompatibility and versatile bio-functionalization towards plasmon-based devices and (bio)molecular sensors.

Nanoparticles Self-Assembly Driven by High Affinity Repeat Protein Pairing.

Proteins are the most specific yet versatile biological self-assembling agents with a rich chemistry. Nevertheless, the design of new proteins with recognition capacities is still in its infancy and has seldom been exploited for the self-assembly of functional inorganic nanoparticles. Here, we report on the protein-directed assembly of gold nanoparticles using purpose-designed artificial repeat proteins having a rigid but modular 3D architecture. αRep protein pairs are selected for their high mutual affinity from a library of 10(9) variants. Their conjugation onto gold nanoparticles drives the massive colloidal assembly of free-standing, one-particle thick films. When the average number of proteins per nanoparticle is lowered, the extent of self-assembly is limited to oligomeric particle clusters. Finally, we demonstrate that the aggregates are reversibly disassembled by an excess of one free protein. Our approach could be optimized for applications in biosensing, cell targeting, or functional nanomaterials engineering.

Assembly of two- and three-dimensionally patterned silicate materials using responsive soft templates.

We report a soft and straightforward method for synthesizing two- and three-dimensionally patterned silicate materials by phase separation using nonionic emulsion templates. Our liquid-state method involves, under controlled atmosphere, the mixing of a condensed silica solution with an oil-in-water emulsion in the presence of a solution of a nonionic emulsifier, Triton X-100. The preparation is stabilized using an organic solvent. The morphology of the silicate materials is significantly modified by changing the reaction conditions or the concentration of the reagents. Three-dimensionally macro and nanoporous continuous films and nanoporous individual spherical particles, both made of amorphous silica, are obtained. The structure of the films and particles is defined by the emulsion template. Films were on average 20 microm thick with a volume-based porosity of approximately 7 x 10(-2) cm(3) g(-1), with pore size correlating well with the size of the oil droplets in the templating emulsion. The siliceous films are bicontinuous leading to large surface areas and openly accessible pores. Individual spheres ranged in size from approximately 1 to 6 microm in diameter with nanoporous openings of 300 nm in diameter. The porosity and integrity of all materials are maintained upon calcination.

Potentiometric determination of the 'formal' hydrolysis ratio of aluminium species in aqueous solutions.

The 'formal' hydrolysis ratio (h = C(OH-)added/C(Al)total) of hydrolysed aluminium-ions is an important parameter required for the exhaustive and quantitative speciation-fractionation of aluminium in aqueous solutions. This paper describes a potentiometric method for determination of the formal hydrolysis ratio based on an automated alkaline titration procedure. The method uses the point of precipitation of aluminium hydroxide as a reference (h = 3.0) in order to calculate the initial formal hydrolysis ratio of hydrolysed aluminium-ion solutions. Several solutions of pure hydrolytic species including aluminium monomers (AlCl3), Al13 polynuclear cluster ([Al13O4(OH)24(H2O)12]7+), Al30 polynuclear cluster ([Al30O8(OH)56(H2O)26]18+) and a suspension of nanoparticulate aluminium hydroxide have been used as 'reference standards' to validate the proposed potentiometric method. Other important variables in the potentiometric determination of the hydrolysis ratio have also been optimised including the concentration of aluminium and the type and strength of alkali (Trizma-base, NH3, NaHCO3, Na2CO3 and KOH). The results of the potentiometric analysis have been cross-verified by quantitative 27Al solution nuclear magnetic resonance (27Al NMR) measurements. The 'formal' hydrolysis ratio of a commercial basic aluminium chloride has been measured as an example of a practical application of the developed technique.