Exploring Nano‐Protein Corona Dynamics: Tracing the Hard‐to‐Soft Corona Transition with Trypsin and Graphene Oxide in a Silver Nanocomposite Model

Abstract In this work, the adsorption of Trypsin on synthesized Silver (Ag), Graphene Oxide: Silver (GO: Ag) nanocomposite (2:1), and Graphene Oxide (GO) nanoparticles (NPs) and, their effect on the conformation of Trypsin was studied by various spectroscopic and microscopic techniques to understand the transition of nano Protein Corona (PC) from “hard” to “soft.” The results showed that the adsorption of Trypsin on synthesized NPs followed the Freundlich adsorption isotherm and pseudo‐second‐order kinetics. The conformational changes that occurred in Trypsin upon interaction with synthesized NPs were investigated by using Circular Dichroism (CD), Fluorescence, and Fourier Transform Infrared (FTIR) spectroscopy. The morphological investigation of nano PC using High‐Resolution Transmission Electron Microscopy (HR‐TEM) and Atomic Force Microscopy (AFM) indicated the formation of hard, semi‐soft, and soft nano PC in the presence of Ag, GO: Ag, and GO NPs, respectively. The negative value of the thermodynamic parameters ΔG , ΔH , and ΔS in all three cases suggests the reaction to be exothermic and spontaneous. The van der Waals interaction and hydrogen bonding are the major forces responsible for binding Trypsin on the surface of Ag, GO: Ag, and GO. The catalytic efficiency of trypsin in the absence and presence of NPs was examined by performing the protein assay showing the highest catalytic activity of pure Trypsin compared with trypsin‐NPs constructs. Our findings provide useful information for the applications of GO‐based nanocomposites for various biological applications.