Surface assembly on biofunctional magnetic nanobeads for the study of protein-ligand interactions.

One of the major challenges of proteomics today is to increase the power potential for the identification of as many proteins as possible and to characterize their interactions with specific free ligands (interactomics) or present on cell walls (cell marker), in order to obtain a global, integrated view of disease processes, cellular processes and networks at the protein level. The work presented here proposes the development of biofunctionalized magnetic nanobeads that might be used for interactomic investigations. The strategy consisted in immobilizing proteins via a non covalent technique that provides greater possibilities for the advent of faster, cheaper and highly miniaturizable protein analysis systems, in particular in situations where the amount of isolated protein is scarce (trace proteins). The advantage of the immobilization technique proposed here over more conventional covalent binding techniques is that it is versatile and universal (not protein specific) thus applicable to a wide range of proteins, in "mild" conditions that are non deleterious to the native structure and bioactivity of the immobilized protein. The feasibility of the technique was investigated using a model protein (streptavidin). The nanobeads were analyzed in size by light diffusion and transmission electronic spectroscopy, and in quantity of immobilized protein using a bioassay developed in the laboratory. Results are promising in that nanobeads exhibited good colloidal stability and surface concentrations in the monolayer range.