A Ready-To-Use Metal-Supported Bilayer Lipid Membrane Biosensor for the Detection of Phenol in Water

This work presents a novel metal-supported bilayer lipid membrane (BLM) biosensor built on tyrosinase to quantitate phenol. The detection strategy is based on the enzyme–analyte initial association and not the commonly adopted monitoring of the redox cascade reactions; such an approach has not been proposed in the literature to date and offers many advantages for environmental monitoring with regard to sensitivity, selectivity, reliability and assay simplicity. The phenol sensor developed herein showed good analytical and operational characteristics: the detection limit (signal-to-noise ratio = 3) was 1.24 pg/mL and the sensitivity was 33.45 nA per pg/mL phenol concentration. The shelf life of the tyrosinase sensor was 12 h and the lifetime (in consecutive assays) was 8 h. The sensor was reversible with bathing at pH 8.5 and could be used for eight assay runs in consecutive assays. The validation in real water samples showed that the sensor could reliably detect 2.5 ppb phenol in tap and river water and 6.1 ppb phenol in lake water, without sample pretreatment. The prospects and applicability of the proposed biosensor and the underlying technology are also discussed.

Supported Bilayer Membranes for Reducing Cell Adhesion in Microfluidic Devices

The high surface area-to-volume ratio of microfluidic channels makes them susceptible to fouling and clogging when used for biological analyses,including cell-based assays. We evaluated the role of electrostatic and van...

Retromer forms low order oligomers on supported lipid bilayers

Retromer is a protein sorting device that orchestrates the selection and export of integral membrane proteins from the endosome via retrograde and plasma membrane recycling pathways. Long standing hypotheses regarding the Retromer sorting mechanism posit that oligomeric interactions between Retromer and associated accessory factors on the endosome membrane drives clustering of Retromer-bound integral membrane cargo prior to its packaging into a nascent transport carrier. To test this hypothesis, we examined interactions between the components of the SNX3-Retromer sorting pathway using quantitative single particle fluorescence microscopy of a reconstituted system comprising a supported bilayer, Retromer, a model cargo protein, the accessory proteins SNX3, RAB7, and the Retromer-binding segment of the WASHC2C subunit of the WASH complex. The predominant species of membrane associated Retromer are low order: monomers (∼18%), dimers (∼35%), trimers (∼24%) and tetramers (∼24%). Unexpectedly, neither cargo nor accessory factors promote Retromer oligomerization on a supported bilayer. The results indicate that Retromer has an intrinsic propensity to form low order oligomers and that neither membrane association nor accessory factors potentiate oligomerization. Hence, Retromer is a minimally concentrative sorting device adapted to bulk membrane trafficking from the endosomal system.

Relevance of charges and polymer mechanical stiffness in the mechanism and kinetics of formation of liponanoparticles probed by the supported bilayer model approach

Parameters controlling the mechanism and kinetics of formation of liponanoparticles are determined using supported lipid bilayer models.