Establishing an AF4-Based Characterization Workflow for the Analysis of Polymeric Nanoparticle-Protein Corona Formation.

2025, January, Volume 1, Issue 1, Page: 21-26

Establishing an AF4-Based Characterization Workflow for the Analysis of Polymeric Nanopart.

Download • 629KB

1. Mr. Manoj Kumar, M. Pharma (Pharmacology) Student, SRK University, Bhopal, MP, India.

2. Dr. Omveer Singh, Associate Professor, MM College of Pharmacy, MM Deemed to be University, Mullana, Haryana.

3. Dr. Girish Vyas, Associate Professor, Faculty of Pharmacology, Career Point University, Kota, Rajasthan, India.

4. Dr. Abhimanu Kumar, Associate Professor, Department of Anatomy, Jakir Hussain Medical College and Research Institute, Raghunathganj, West-Bengal, India.

5. Dr. Shailesh Sharma, Research Director & Principal, Faculty of Pharmacy, Dr. KN Modi University, Rajasthan, India.

6. Dr. Hansraj Bishnoi, Principal, Khatu Shyam School of Pharmacy, Rajasthan.

Corresponding Author:

Mr. Manoj Kumar

M.Pharma (Pharmacology) Student

SRK University, Bhopal, MP

Abstract:

Accurate characterization of nanoparticle-protein complexes, particularly the protein corona formed in biological media, is crucial for understanding their behavior and potential applications. Conventional centrifugation-wash methods, however, introduce artifacts by disrupting the delicate equilibrium of these complexes. This study explores asymmetric flow field-flow fractionation (AF4) as a non-disruptive alternative for analyzing nanoparticle-protein interactions under physiologically relevant conditions.1 Model surface-modified polystyrene latex nanoparticles were incubated in protein-containing media, simulating cell culture conditions, and subsequently analyzed using AF4 with in-line detectors. The results demonstrate that AF4 enables high-resolution separation and characterization of intact nanoparticle-protein complexes, preserving their native state. This approach avoids the limitations of centrifugation-wash methods, which can alter nanoparticle parameters and protein corona composition. By offering a robust and harmonized pipeline for in-situ characterization, AF4 facilitates a more accurate and biologically relevant understanding of nanoparticle-protein interactions.