A Comparative Study on Surface Treatments in the Immobilization Improvement of Hexahistidine-tagged Protein on the Indium Tin Oxide Surface
- Corresponding Author:
- Manel Ben Ismail
Molecular Biology and Biochemistry Research Center for Nanomedicine (Cibbim-Nanomedicine)
Vall d’Hebron Institut de Recerca (VHIR). Passeig de la Vall d’Hebron
11912908035 Barcelona, Spain
E-mail: [email protected]
Received Date: April 11, 2016; Accepted Date: April 24, 2016; Published Date: April 30, 2016
Citation: Ismail MB, Pastor NC, Soler EP, Soltani A, Othmane A (2016) A Comparative Study on Surface Treatments in the Immobilization Improvement of Hexahistidine-tagged Protein on the Indium Tin Oxide Surface. J Nanomed Nanotechnol 7: 372. doi:10.4172/2157-7439.1000372
Copyright: © 2016 Ismail MB, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Biosensors are becoming increasingly important in various sectors. However, choosing transducer type and surface treatment still faces severe limitations. Indium-tin oxide is widely used in electrochemical sensing, though surface cleaning and functionalization are not always straightforward and often poorly controlled. Here, we have covalently immobilized a hexahistidine-tagged model protein on three different treated ITO surfaces using a Ni2+ chelator moiety. The study shows that compared to two other treatments (H2O/H2O2/NH4OH and KOH treatments), the ITO Piranha treatment gives satisfactory results in term of yields of EC12 protein immobilized on the surface. The study in detail of treatments effect was realized by applying a combination of local and global techniques such X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), Attenuated total reflectance Fourier transform infrared spectroscopy (FTIR-ATR), contact angle measurements, and electrochemical impedance spectroscopy (EIS). The biofunctionalization allowed proteins to move freely around their anchoring bond. As the polyhistidine tags are widely used in protein engineering, such substrates offer a large panel of applications. This covalent and oriented immobilization process of recombinant proteins applied to ITO could provide transparent electroactive surfaces of high quality for electrochemical detection of soluble compounds, as well as cells.