O. A. Lytkina, A. A. Sazonov
Siberian State Medical University, Researcher of Central Research Laboratory, Russia
The development of hybrid magnetic nanobiostructures, which are able to bind to target biomolecule in a specific way, opens some attractive opportunities in the area of highsensitivity diagnostic systems de velopment .
The goal of this research was to examine the functional activity of the proteins immobilized on nano particles surface. The carbon coated iron nanoparticles served as a core of the developed construction . Bovine serum albumin, interferon alpha and horseradish peroxidase were used as model biomolecules. For covalent immobilization of proteins the surface of nanoparticles was modified with original diazonium salt –
4 carboxy benzenediazonium tosylate . The proteins were covalently bound to nanoparticles by carbo diimide activation. The nanohybryd constructions received as the result, were separated from the mixture by
magnetic precipitation and centrifugation and washed with phosphate buffer and water at least four times.
The proteinnanoparticles complex formation was confirmed by FTIR spectroscopy (in IRspectra of complexes characteristic absorption bands of protein appeared in the region of 16001500 cm1). Spectropho tometric analysis has shown that proteinbinding capacity of nanoparticles constituted 0,10,4*106 Mol per gramm. It was determined, that horseradish peroxidase partially retained the specific enzymatic activity after immobilization. The capability of nanoparticles containing interferon alpha to bind with the specific antibo dies was demonstrated by ELISA. Therefore the immobilization doesn’t lead to the distortion of the protein molecule spatial structure.
There was developed the fundamental approach to the creation of the hybrid bionanocomposite con structions which include the magnetic nanoparticles covalently bound to proteins. This approach constitutes the main prerequisite for the development of new generation effective magnetosensitive biosensors.
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Материал взят из: Науки о человеке: Сборник статей по материалам XII Российского конгресса молодых ученых с международным участием (Томск, 26-27 мая 2011 г.)