Effets des solutions tampons sur les propriétés adsorbantes d'une résine fonctionnalisée vis-à-vis de l’HSA et de l’IgG humaines

Abstract

Medicaments derived from plasma represent a particular sector of pharmaceutical field. They are produced by fractionation of crude material, human plasma, allowing the isolation of proteins of interest. These proteins highly characterized are indispensable to treat several pathologies and thus save lives. Plasmatic fractionation consists to prepare these therapeutic products using precis and performant technics leading to the best yield both on quantity and quality. Among these technics, we attempted to integrate the affinity chromatography method, IMAC. A new functionalized chromatoghraphic adsorbant is used [N,N-bis(salicylidenepropylenetriamine)-aminomethyl polystyrène] and which can fix different metals as Cu(II). The aim of this study is to find the most adequate conditions for the adsorption and desorption of specific proteins on resin. The purity of obtained effluent was controlled using SDS-PAGE. Results showed that the resin-Cu (II) was capable to purify a quantity of pure HSA using the elution buffer (5 mM Tris-HCl, 10 mM imidazole, 10 mM MOPS and 35 mM NaCl) and also the buffer (10 mM buffer phosphate, 35 mM NaCl and 5 mM imidzole). Moreover, a quantity of pure IgG was recuperated adding a low concentration of imidazole (5 or 10 mM) to the elution buffer (10 mM buffer phosphate and 5 mM MOPS). In conclusion, this resin is a good chelator matrix of bivalent metals and may be used to purify IgG and HAS from human plasma for the diagnostic and/or therapeutic purposes.