Electrochemical Characterization of Self-Assembled Monolayers on Gold Substrates Derived from Thermal Decomposition of Monolayer-Protected Cluster Films

Networked films of monolayer-protected clusters (MPCs), alkanethiolate-stabilized gold nanoparticles, can be thermally decomposed to form stable gold on glass substrates that are subsequently modified with self-assembled monolayers (SAMs) for use as modified electrodes. Electrochemical assessment of these SAM-modified gold substrates, including double-layer capacitance measurements, linear sweep desorption of the alkanethiolates, and diffusional redox probing, all show that SAMs formed on gold supports formed from thermolysis of MPC films possess substantially higher defect density compared to SAMs formed on traditional evaporated gold. The density of defects in the SAMs on thermolyzed gold is directly related to the strategies used to assemble the MPC film prior to thermolysis. Specifically, gold substrates formed from thermally decomposing MPC films formed with electrostatic bridges between carboxylic acid-modified MPCs and metal ion linkers are particularly sensitive to the degree of metal exposure during the assembly process. While specific metal dependence was observed, metal concentration within the MPC precursor film was determined to be a more significant factor. Specific MPC film linking strategies and pretreatment methods that emphasized lower metal exposure resulted in gold films that supported SAMs of lower defect density. The defect density of a SAM-modified electrode is shown to be critical in certain electrochemical experiments such as protein monolayer electrochemistry of adsorbed cytochrome c. While the thermal decomposition of nanoparticle film assemblies remains a viable and interesting technique for coating both flat and irregular shaped substrates, this study provides electrochemical assessment tools and tactics for determining and controlling SAM defect density on this type of gold structure, a property critical to their effective use in subsequent electrochemical applications

The contribution of X-linked coding variation to severe developmental disorders

Abstract: Over 130 X-linked genes have been robustly associated with developmental disorders, and X-linked causes have been hypothesised to underlie the higher developmental disorder rates in males. Here, we evaluate the burden of X-linked coding variation in 11,044 developmental disorder patients, and find a similar rate of X-linked causes in males and females (6.0% and 6.9%, respectively), indicating that such variants do not account for the 1.4-fold male bias. We develop an improved strategy to detect X-linked developmental disorders and identify 23 significant genes, all of which were previously known, consistent with our inference that the vast majority of the X-linked burden is in known developmental disorder-associated genes. Importantly, we estimate that, in male probands, only 13% of inherited rare missense variants in known developmental disorder-associated genes are likely to be pathogenic. Our results demonstrate that statistical analysis of large datasets can refine our understanding of modes of inheritance for individual X-linked disorders

Predicting the Loading of Virus-Like Particles with Fluorescent Proteins

The virus-like particle (VLP) of the Cowpea Chlorotic Mottle Virus (CCMV) has often been used to encapsulate foreign cargo. Here we show two different rational design approaches, covalent and noncovalent, for loading teal fluorescent proteins (TFP) into the VLP. The covalent loading approach allows us to gain control over capsid loading on a molecular level. The achieved loading control is used to accurately predict the loading of cargo into CCMV VLP. The effects of molecular confinement were compared for the differently loaded VLPs created with the covalent method. We see that the loading of more than 10 fluorescent proteins in the 18 nm internal cavity of the CCMV capsid gives rise to a maximum efficiency of homo-FRET between the loaded proteins, as measured by fluorescence anisotropy. This shows that already at low levels of VLP loading molecular crowding starts to play a role