Logan Mackay

Gel free analysis of the proteome of intracellular Leishmania mexicana

Investigating the proteome of intracellular Leishmania amastigotes has recently become possible due to the exploitation of fluorescence activated intracellular parasite sorting. Here, we employed this technology in combination with gel free analysis to greatly improve proteome coverage and suggest proteins putatively secreted by the parasites. In total, 1764 proteins were identified of which 741 had not been reported before. Protein abundance indices were calculated to rank individual proteins according to their abundance in vivo. Using the LeishCyc resource, an overview of metabolically relevant proteins was produced that integrated protein abundance data. Bioinformatic analysis identified 143 proteins possibly secreted by L. mexicana amastigotes, half of which have no known function. The data provide a useful resource, e.g. for modelling metabolic flux or selecting novel vaccine antigens.

Copper Catalysis in Living Systems and In Situ Drug Synthesis

The copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction has proven to be a pivotal advance in chemical ligation strategies with applications ranging from polymer fabrication to bioconjugation. However, application in vivo has been limited by the inherent toxicity of the copper catalyst. Herein, we report the application of heterogeneous copper catalysts in azide-alkyne cycloaddition processes in biological systems ranging from cells to zebrafish, with reactions spanning from fluorophore activation to the first reported in situ generation of a triazole-containing anticancer agent from two benign components, opening up many new avenues of exploration for CuAAC chemistry.

Test structures and a measurement system for characterising the lifetime of EWOD devices

This paper presents a methodology together with a characterisation system for rapidly and quantitatively evaluating the lifetime and reliability of Electro-Wetting-on-Dielectric (EWOD) microfluidic devices. By studying the contact angle (CA) change when electrowetting forces are applied on the test structure, the number of times of a droplet can be switched between relaxed and actuated states can be characterised. This paper describes the development of an automated measurement system together with test structures designed to quickly characterise and optimise EWOD dielectric layer compositions. This enables the rapid determination of the driving voltage that results in the longest lifetime of the device, providing the tools to optimise both the process, architecture and the voltage driving scheme.