Development of native comprehensive three-dimensional liquid chromatography-mass spectrometry for unravelling biomolecular interaction networks in support of post-genomic biotechnology in healthcare: 3DbioNET (FWOAL866)

The possibility to identify biomolecular interactions involved in cell regulation provides new opportunities to detect and treat diseases. The function of biomolecules should not only be understood for isolated proteins but also in the context of other biomolecules, since > 80% of proteins exert function as part of larger assemblies. Despite their importance, there are no high-throughput techniques that allow to analyze biomolecular interactions directly in complex matrices.

This project aims at realizing novel separation technology, allowing for unprecedented liquid chromatography (LC) separations, and application to profile interaction networks directly in contemporary life-science mixtures. New technological innovations that will be established include the development of functionalized polymer-monolithic stationary phases allowing for native high-resolution separations at low salt content, the development of microfluidic modulator chips for sample transfer between columns in comprehensive three-dimensional LC (LCxLCxLC) and, prototyping of a multi-nozzle nano-electrospray device allowing for sub-10 nL/min flow systems to maintain macromolecular assemblies intact during infusion in the mass spectrometer. Detection of proteins involved in interaction networks, targeting biomarker discovery, will be performed directly in contemporary life-science mixtures via profiling of retention-time differences in the presence of protein binding ligands.

Keywords:liquid, chromatography

Disciplines:Clinical chemistry