Technical Report on the reduced graphene oxide biosynthesis protocol for biological applications

The interest of scientific community on carbon-based smart materials is growing and, especially focus on graphene oxide (GO) and reduced graphene oxide (rGO). An increasing number of bio-applications such as biological applications as bacterial inhibition, drug delivery and photothermal therapy aims the use of GO and rGO. For this reason, the methods used for the synthesis of graphene materials are more important because same of those procedures imply chemical reactions that involve hazardous and toxic reagents. In fact, the biocompatibility and toxicological activity of graphene-related materials is related to the methodologies employed for the synthesis that determine the carbon/oxygen (C/O) ratio of graphene oxide species. In this technical report, we focused on the synthesis of GO by means of that lead to a biocompatible GO form with a lower oxygen content. Thus, the synthesis of rAsGFP-rGO with the green fluorescent protein allowed us to obtain a biocompatible materials, without using hazardous and toxic reagents. This biocompatibility is the most important prerogative for the use of GO in biological activity assays as reported in several publications.