Hydrophilic Submicron Nanogel Particles for Specific Recombinant Proteins Extraction and Purification

摘要

In biomedical diagnosis and bionanotechnologies, the extraction and purification of proteins and protein derivatives are of great interest. In fact, to purify recombinant proteins for instance, new methodologies and well appropriate material supports need to be established and also to be evaluated. In this work, hydrophilic nanohydrogel particles were prepared for recombinant proteins extraction for purification purpose. The prepared nanohydrogel polymer-based particles are hydrophilic below the volume phase transition temperature (TVPT) and dehydrated above the TVPT, due to the thermally sensitive poly(N-alkyl acrylamide) and poly(N-alkyl methacrylamide) derivatives. Then, the use of heavy metal ions in the presence of such functional particles should specifically capture recombinant proteins (i.e., proteins bearing a poly(histidine) part). In order to understand and to optimize the specific capture and the purification of recombinant proteins, various parameters have been investigated as a systematic study. Firstly, the adsorption was investigated as a function of pH and protein concentration. According to high hydration of the prepared nanohydrogel, no marked adsorption was observed. Secondly, the effect of pH was investigated and found to be the driven parameter affecting the metal ions immobilization and the recombinant proteins complexation. As a result, high protein complexation was observed at basic pH compared to non-complexation at acidic pH medium. The immobilized proteins via complexation were released by changing the pH. This decomplexation seems to be effective but depends on fixation conditions and particle surface structure.