PACS 07.60.Hv, 78.30.Jv, 85.60.Me, 87.80.+s

A biosensor approach to probe the structure and function of the adsorbed proteins: fibrinogen at the gold surface

B. A. Snopok*, K. V. Kostyukevych, O. V. Rengevych,
Y. M. Shirshov, E. F. Venger
Institute of Semiconductor Physics, NAS Ukraine, 45 prospekt Nauki, Kyiv, 252028, Ukraine
* B.A.S.: Email: snopok@isp.kiev.ua; Telephone: +380 (44) 265 56 26; Phone (Voice&Fax): +380 (44) 265 18 27.

I. N. Kolesnikova, E. V. Lugovskoi
Palladin Institute of Biochemistry, NAS Ukraine, ul. Leontovicha 9, Kyiv-030, 252030 Ukraine

Abstract. The kinetics of adsorption and surface structure of adsorbed layers of the human fibrinogen on the gold surface, determined by Surface Plasmon Resonance (SPR) and Atomic Force Microscopy (AFM) analysis, was employed to probe the lateral distribution and preferred orientation of protein molecules within the monolayer. In this study, special sets of immunoassays are presented for fibrinogen adsorption/conformation analysis. The results show that kinetic parameters of antigen-antibody interactions are directly related to the interfacial conformation of fibrinogen molecules. Various interfacial structures of adsorbed fibrinogen aggregates, namely single, bi- and three- molecular aggregates, were obtained using a combination of AFM imaging and SPR analysis. Adsorption of fibrinogen onto the surface of polycrystalline gold is a complex process including surface-induced unfolding, local self-assembly and adsorption, occurring concurrently with and on the time scale of each other. This result confirmed the utility of the proposed approach for detecting the spatial distribution and biofunctional properties of specific proteins adsorbed from biological liquids in biosensors.

Keywords: surface plasmon resonance, adsorbed proteins, fibrinogen, DD-fragment, E-fragment, biosensor, monoclonal antibodies, AFM, film chemistry control, polycrystalline gold films.

Paper received 10.10.98; revised manuscript received 10.10.98; accepted for publication 28.10.98.

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