An elastomeric stamp, containing defined features on the micrometer scale, was used to imprint gold surfaces with specific patterns of self-assembled monolayers of alkanethiols and, thereby, to create islands of defined shape and size that support extracellular matrix protein adsorption and cell attachment. Through this technique, it was possible to place cells in predetermined locations and arrays, separated by defined distances, and to dictate their shape. Limiting the degree of cell extension provided control over cell growth and protein secretion. This method is experimentally simple and highly adaptable. It should be useful for applications in biotechnology that require analysis of individual cells cultured at high density or repeated access to cells placed in specified locations.

This paper reports a simple, one-pot, template-free synthesis of flower-like Au nanoparticles (three-dimensional branched nanoparticles with more than 10 tips) with high yield and good size monodispersity at room temperature. The size of the Au nanoflowers could be tuned by controlling the composition of the starting reaction mixture. The key synthesis strategy was to use a common Good's buffer, HEPES, as a weak reducing and particle stabilizing agent to confine the growth of the Au nanocrystals in the special reaction region of limited ligand protection (LLP). Time-course measurements by UV-vis spectroscopy and TEM were used to follow the reaction progress and the evolution of the flower-like shape. The Au nanoflowers exhibited strong surface-enhanced effects which were utilized in the design of an efficient, stable, and nontoxic Raman-active tag for in vivo applications.

In this work, three-dimensional branched gold nanocrystals were produced at high yield by reacting an aqueous solution of chloroauric acid with a Good's buffer, HEPES, at room temperature. This particular method of preparation was scalable to gram-quantity. The branched nanocrystals containing one to eight tips were stable at room temperature and could be stored as a powder after freeze-drying. They were, however, unstable at higher temperatures and transformed into spherical particles upon boiling. The formation of the branched gold nanocrystals was kinetically controlled, as shown by the dependence of shapes on temperature and precursor salt concentration. The growth of branched gold nanocrystals in the HEPES buffer was monitored by microscopic and spectroscopic techniques, allowing the detection of several key intermediates in the growth process. Piperazine in HEPES molecule was identified as the principal moiety responsible for forming highly branched Au nanocrystals in the HEPES buffer.

This paper describes the synthesis of single-crystalline Ag nanoplates using the extract of unicellular green alga Chlorella vulgaris at room temperature. Proteins in the extract were involved in the biological synthesis, providing the dual function of Ag ion reduction and shape-controlled synthesis of nanosilver. Hydroxyl groups in Tyr residues and carboxyl groups in Asp and/or Glu residues were further identified as the most active functional groups for Ag ion reduction and for directing the anisotropic growth of Ag nanoplates, respectively. The kinetics of Ag ion reduction in biological systems was discussed and probed by using custom-designed peptides. The results showed the Tyr content (the reduction source) and the content of Ag complexers (the reaction inhibitors, e.g., His and Cys) in the protein molecules as important factors affecting the reduction kinetics. The comprehensive system identification effort has led to the design of a simple bifunctional tripeptide (DDY-OMe) with one Tyr residue as the reduction source and two carboxyl groups in the Asp residues as shape-directors, which could produce small Ag nanoplates with low polydispersivity in good yield (>55%). The roles of the carboxyl groups in the formation of Ag nanoplates were also discussed.

Peptides are multifunctional reagents (reducing and capping agents) that can be used for the synthesis of biocompatible metal nanoparticles under relatively mild conditions. However, the progress in peptide synthesis of metal nanoparticles has been slow due to the lack of peptide design rules. It is difficult to establish sequence-reactivity relationships from peptides isolated from biological sources (e.g., biomineralizing organisms) or selected by combinatorial display libraries because of their widely varying compositions and structures. The abundance of random and inactive amino acid sequences in the peptides also increases the difficulty in knowledge extraction. In this study, a "bottom-up" approach was used to formulate a set of rudimentary rules for the size- and shape-controlled peptide synthesis of gold nanoparticles from the properties of the 20 natural alpha-amino acids for AuCl(4)(-) reduction and binding to Au(0). It was discovered that the reduction capability of a peptide depends on the presence of certain reducing amino acid residues, whose activity may be regulated by neighboring residues with different Au(0) binding strengths. Another finding is the effect of peptide net charge on the nucleation and growth of the Au nanoparticles. On the basis of these understandings, several multifunctional peptides were designed to synthesize gold nanoparticles in different morphologies (nanospheres and nanoplates) and with sizes tunable by the strategic placement of selected amino acid residues in the peptide sequence. The methodology presented here and the findings are useful for establishing the scientific basis for the rational design of peptides for the synthesis of metal nanostructures.

The clustering and stability of magnetic nanoparticles coated with random copolymers of acrylic acid, styrenesulfonic acid, and vinylsulfonic acid has been studied. Clusters larger than 50 nm are formed when the coatings are made using too low or too high molecular weight polymers or using insufficient amounts of polymer. Low-molecular-weight polymers result in thin coatings that do not sufficiently screen van der Waals attractive forces, while high-molecular-weight polymers bridge between particles, and insufficient polymer results in bare patches on the magnetite surface. The stability of the resulting clusters is poor, but when an insufficient polymer is used as primary coating, and a secondary polymer is added to coat remaining bare magnetite, the clusters are stable in high salt concentrations (>5 M NaCl), while retaining the necessary cluster size for efficient magnetic recovery. The magnetite cores were characterized by TEM and vibrating sample magnetometry, while the clusters were characterized by dynamic light scattering. The clustering and stability are interpreted in terms of the particle-particle interaction forces, and the optimal polymer size can be predicted well on the basis of these forces and the solution structure and hydrophobicity of the polymer. The size of aggregates formed by limited polymer can be predicted with a diffusion-limited colloidal aggregation model modified with a sticking probability based on fractional coating of the magnetite cores.

In this work, single-crystalline gold nanoplates were produced by treating an aqueous solution of chloroauric acid with the extract of the unicellular green alga Chlorella vulgaris at room temperature. The results suggest proteins as the primary biomolecules involved in providing the dual function of Au(III) reduction and the size- and shape-controlled synthesis of the nanogold crystals. A protein with a molecular weight of approximately 28 kDa was isolated and purified by reversed-phase HPLC; this protein tested positive for the reduction of chloroauric acid in aqueous solution. The isolated protein (named gold shape-directing protein, or GSP for convenience) was then used to produce gold nanoplates with distinctive triangular and hexagonal shapes in high yields (approximately 90 %). The kinetics of the reduction reaction could be manipulated through changes in the GSP concentration to produce plates with lateral sizes ranging from nanometers to micrometers. The growth of gold nanoplates in the GSP solution with time was monitored by microscopic and spectroscopic techniques, thereby allowing the detection of several key intermediates in the growth process.

The amino acid arginine is frequently used as a solution additive to stabilize proteins against aggregation, especially in the process of protein refolding. Despite arginine's prevalence, the mechanism by which it stabilizes proteins is not presently understood. We propose that arginine deters aggregation by slowing protein-protein association reactions, with only a small concomitant effect on protein folding. The associated rate effect was observed experimentally in association of globular proteins (insulin and a monoclonal anti-insulin) and in refolding of carbonic anhydrase. We suggest that this effect arises because arginine is preferentially excluded from protein-protein encounter complexes but not from dissociated protein molecules. Such an effect is predicted by our gap effect theory [Baynes and Trout (2004) Biophys. J. 87, 1631] for "neutral crowder" additives such as arginine which are significantly larger than water but have only a small effect on the free energies of isolated protein molecules. The effect of arginine on refolding of carbonic anhydrase was also shown to be consistent with this hypothesis.