Obtaining protein crystals suitable for X-rays scattering is requested to resolve protein functionalities, which is one of the most important goal of biological research. In this domain, microfluidics has recently proved be a powerful, rapid and efficient approach: by rapidly screening an important number of different conditions, and exploiting scale reduction, accelerates by orders of magnitude the determination of optimal grow conditions. This approach led to crytallise for the first time integrin.
The aim of PROTEMICROFLU is to develop a new microfluidic approach to protein crystallisation. The system we propose is an array of evaporation microchambers, moulded on a microfluidic chip, filled by solutes of different concentration and exposed, through a membrane, to air fluxes driven through microchannels, placed at a second level.
The system is made by using multi layer lithography, a technology well handled by the host laboratory. The agents will be concentrated by evaporation of the solvent through the membranes, achieving supersaturated states which eventually crystallize. In a second step, the system will be equipped by microvalves to facilitate fluid handling and favour integration and compactness.
The main goal is to map the protein phase diagram and look for optimal crystallization conditions; owing to the flexibility and speed of the handling of the working fluids, it will be possible to interactively optimize crystal sizes and structures, along with understanding the whole kinetic process. Compared to existing microfluidic systems, the interactivity and improved control represent substantial progress.
The study will first address standard proteins (lysozyme and g crystallin) and will aim at considering proteins unresolved by traditional techniques. The project feasibly is demonstrated by preliminary studies; moreover, the host laboratory expertise (microtechnology) will provide a complementary formation to the candidate background (protein interaction).
Fields of science
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