Electrochemical desorption of fibrinogen from gold

Abstract

The electrochemically induced desorption of Oregon green labeled fibrinogen layers from clean gold surfaces at negative potentials has been probed using capacitance, fluorescence microscopy, and atomic force microscopy. Capacitance measurements on fibrinogen layers indicate that desorption occurs at potentials more negative than -0.8 V and that complete desorption occurs when the electrode is biased at -1.2 V. Significantly, the fluorescence intensity initially increases as the dye labeled protein is electrochemically desorbed due to a decrease in quenching by the gold surface. Following this initial increase, the protein diffuses into solution and the fluorescence intensity decreases over time. More than 90% of the dye labeled fibrinogen is desorbed and diffuses out of the confocal volume in less than 2000 s when the potential is stepped to -1.2 V. AFM before and after application of the desorbing potential confirms removal of the protein. Collection of the desorbed protein in solution reveals a surface coverage of (4.0 +/- 2.3) x 10(-13) mol cm(-2) or an area of occupation of 400 +/- 140 nm(2) per molecule, which indicates that the protein is not extensively spread on the bare gold surface. Significantly, SDS-PAGE analysis indicates that the adsorption-desorption cycle dramatically effects the protein structure, with the electrochemically desorbed fibrinogen showing extensive fragmentation compared to native protein.