Advancing Protein Corona Analysis of Graphene Oxide

This study addresses limitations in standard SDS-based protocols for protein corona analysis of graphene oxide (GO), a nanomaterial characterized by strong and heterogeneous protein interactions. Conventional elution methods failed to achieve complete protein desorption from GO surfaces, leading to underrepresentation of bound biomolecules. To overcome this, a modified stepwise extraction protocol incorporating chaotropic agents (urea and thiourea) was developed, enabling near-complete protein recovery. SDS-PAGE, densitometric analysis, and mass spectrometry demonstrated improved total protein yield and enhanced detection of more hydrophobic proteins compared to the standard protocol. These findings highlight the need for tailored extraction strategies to accurately characterize the biomolecular corona of advanced nanomaterials, with important implications for nanosafety assessment and reliable evaluation of GO bioactivity.