Immobilization of histidine-tagged proteins on gold surfaces using chelator thioalkanes

The reversible, oriented immobilization of proteins on solid surfaces is a prerequisite for the investigation of mol. interactions and the controlled formation of supramol. assemblies. This paper describes a generally applicable method using a synthetic chelator thioalkane that can be self-assembled on gold surfaces. The reversible binding of an anti-lysozyme Fab fragment modified with a C-terminal hexahistidine extension was monitored and the apparent dissocn. consts. detd. using surface plasmon resonance. Infra-red spectroscopy demonstrated that the secondary structure of the protein was unaffected by the immobilization process. The retention of functionality of the immobilized protein was also successfully demonstrated. Given the mild reaction conditions and reversibility, this method of oriented immobilization of proteins opens possibilities for the development of biosensors. [on SciFinder (R)]