Optimization of the interaction of graphene quantum dots with lipase for biological applications

Graphene quantum dots (GQDs) are known as emerging sub-10 nm nanoparticles (NPs), which are in fact few-layered pieces of graphene, capable of emitting blue fluorescence, when exposed to 360 nm UV light. Understanding the details of the interaction between GQDs and lipase can serve as a critical step for improving the biological outcome of GQD-derived drug-delivery and diagnosis systems. The interaction occurs in the form of surface adsorption, which can subsequently influence the physicochemical properties of both the NP and the protein. Hence, a systematic approach was taken here to optimize the GQDs' synthesis conditions in order to achieve the highest possible quantum yield (QY). Furthermore, to understands the influence of the interaction of GQDs and lipase, on both the activity of lipase and the emission intensity of GQDs, various incubation conditions were tested to achieve optimized conditions over central composite design algorithm by Design-Expert®, using response surface methodology. The results show that the GQDs fabricated by thermal decomposition of citric acid at 160°C, with a heating duration of 55 min, obtain almost three times higher QY than the highest values reported previously. The best enzymatic activity after the formation of the hard corona, as well as the highest fluorescent emission, were achieved at GQD-to-enzyme ratios within the rage of 23-25%, at temperatures between 41 and 42°C, for 6-8 min. In the aforementioned condition, the enzyme retains 91-95% of its activity and the NP preserves about 80-82% of its fluorescence intensity after incubation.