Immobilization of cytochrome c and its application as electrochemical biosensors

Cytochrome c (Cyt c) has been used as a model protein to investigate the characters of modified electrodes by many researchers. It has been also employed to construct biosensors to detect hydrogen peroxide, nitrate, superoxide, and etc. Cyt c immobilization techniques, including physical adsorption, entrapment in hydrogel or polymers, layer-by-layer assembly, Langmuir-Blodgett, and covalent attachment are discussed followed by various electrochemical methods applied in the electrode modification. The exploration of some modified protein electrodes, for example, screen printed, microperoxidase and engineered Cyt c are also presented. The preparation, characterizations and some properties of nanocomposites to modify electrode surface for immobilizing Cyt c are highlighted. This review is attempted to discuss the influences of the physical and chemical properties of the substrate materials, such as specific area and surface charge on the protein loading and electron transfer of Cyt c briefly. The comparative information of Cyt c-based electrochemical modified electrodes, such as average surface coverage, sensitivity, linear range, and detection limit of the analyte of interest is also summarized.