Micro and nano particles in biotechnology

Research project "Micro and Nano Particles in Biotechnology" is composed of two segments that play a central role in development of modern bio-pharmaceuticals. Common to both parts are knowledge and technology development for preparation and characterization of protein micro and nano particles applicable in modern biotechnology. The first part involves intracellular micro-aggregation of proteins leading to inclusion body formation. According to our hypothesis (already partially confirmed) that in some properly folded proteins (e.g. TNF-alpha) micro-aggregation and inclusion body formation can be controlled by modulation of hydrophobicity/hydrophylicity of the affinity oligo-histidine tag. We propose a novel function and applicability of histidine tags, primarily used for easier protein isolation. During biosynthesis inclusion bodies will serve for protein storage with the aim of directing and controlling micro-aggregation and precipitation of biologically active proteins, but not improperly folded ones. TNF-alpha will be the main oligomeric protein model and GFP a monomeric, both with histidine tags attached to the N-termini. Design of novel "hybrid" histidine tags for above described purposes is also anticipated. The second part deals with preparation of macro-molecular aggregates (nano particles) of highly purified recombinant proteins as a new approach to pharmaceutical formulations. An original way of using controlled and reversible micro-aggregation of histidine-tagged molecules upon metal ion addition will be applied. This should further result in development of protein formulations with sustained release and higher in vivo stability.