Technical tip: high-resolution isolation of nanoparticle-protein corona complexes from physiological fluids

Area of Science:

• Nanomedicine
• Biomaterials Science
• Analytical Chemistry

Background:

• Nanoparticles (NPs) interact with biological fluids, forming a protein corona that dictates their biological identity.
• Understanding the protein corona is crucial for predicting nanoparticle behavior in vitro and in vivo.
• Current methods for NP-protein corona isolation can alter their structure and composition.

Purpose of the Study:

• To develop a one-step methodology for recovering NPs from biological media.
• To preserve the native structure and composition of the NP protein corona during isolation.
• To enable accurate characterization of NP-protein complexes for biological studies.

Main Methods:

• Incubation of diverse nanoparticles in biological fluids.
• Isolation of NPs and their protein corona using sucrose gradient ultracentrifugation.
• Characterization of isolated complexes via Dynamic Light Scattering, Nanoparticle Tracking Analysis, SDS-PAGE, and LC-MS.

Main Results:

• A novel, one-step method successfully recovers NPs from complex biological fluids.
• The proposed methodology preserves the NP protein corona's structure and composition.
• This technique offers higher resolution and lower impact compared to conventional centrifugal approaches.

Conclusions:

• The developed protocol provides a versatile and effective means for isolating NP-protein complexes.
• This method facilitates accurate analysis of nanoparticle biological identity.
• It is applicable to various nanomaterials and biological fluids, advancing nanomedicine research.