Protein Corona Modulates Extracellular Vesicle Recognition at Glycan-Based Nanobiointerfaces

Area of Science:

• Biomaterials Science
• Nanotechnology
• Extracellular Vesicle Biology

Background:

• Precise surface engineering is crucial for characterizing extracellular vesicles (EVs) and their biological interactions.
• Gold substrates modified with self-assembled monolayers (SAMs) and gold nanoparticles (AuNPs) enhance surface area and functionality for nanostructure assembly.

Purpose of the Study:

• To develop and characterize novel glycan-based nanobiointerfaces for studying extracellular vesicle (EV) interactions.
• To investigate the impact of protein corona removal on EV surface marker accessibility and recognition.
• To differentiate malignant and non-malignant EVs based on their binding behavior to glycan interfaces.

Main Methods:

• Fabrication of gold substrates with C6 SAMs, AuNPs, and layer-by-layer assembly of glycan nanostructures.
• Electrochemical methods and atomic force microscopy (AFM) for interface monitoring.
• Affinity isolation, preconcentration, and protein corona removal of EVs (MDA-MB-231 and RWPE-1).
• ELISA for evaluating antibody-mediated recognition post-corona removal.
• Surface Plasmon Resonance (SPR) with a physics-informed neural network for analyzing EV-glycan interactions.

Main Results:

• Optimized SAMs and AuNP deposition created robust nanobiointerfaces.
• Protein corona removal significantly improved EV surface marker accessibility for antibody recognition.
• SPR analysis revealed distinct binding phenotypes of malignant EVs compared to non-malignant EVs on glycan surfaces.
• The physics-informed neural network model effectively reduced bulk refractive index interference in SPR data.

Conclusions:

• Glycan-based nanobiointerfaces enable sensitive characterization of EV binding.
• Protein corona composition and glycan interactions are critical factors in EV recognition.
• These findings support the potential of glycan-based nanobiointerfaces for advancing extracellular vesicle research and liquid biopsy development.