Ni(ii)-modified solid substrates as a platform to adsorb His-tag proteins

Abstract

This work investigated a simple and versatile modification to a solid substrate to develop electrochemical bio-recognition platforms based on bio-affinity interactions between histidine (His)-tagged proteins and Ni(ii) surface sites. Carboxylate (COO)-functionalized substrates were prepared in multiple steps, initiated with an amino-terminated self-assembled monolayer (SAM) on polycrystalline gold. Surface enhanced Raman spectroscopy (SERS), quartz crystal microbalance with dissipation monitoring (QCM-D) and contact angle measurements were used to follow the modification process. Upon completion of the modification process, the surface COO-Ni(ii) chelate complex and the coordination mode used to bind the His-tag proteins were characterized by X-ray absorption near-edge spectroscopy (XANES). Finally, the electrochemical stability and response of the modified substrates were evaluated. The versatility of the modification process was verified using silica as the substrate. QCM-D and SERS results indicated that two types of films were formed: a COO-terminated SAM, which resulted from the reduction of previously incorporated surface aldehyde groups, and a physically adsorbed polymeric glutaraldehyde film, which was produced in the alkaline medium. XANES spectral features indicated that COO-Ni(ii) formed a non-distorted octahedral complex on the substrate. The electrochemical stability and response towards a redox mediator of the COO-Ni(ii)-terminated SAM indicated that this platform could be easily coupled to an electrochemical method to detect bio-recognition events. © 2013 The Royal Society of Chemistry.