<p>Accurate and robust proteomics characterization of the protein corona is crucial for both diagnostic and therapeutic applications of nanomedicine technologies. This Perspective focuses on mass spectrometry-based characterization of the protein corona, emphasizing its role in identifying and quantifying the proteins associated with it. The discussion encompasses three major steps: protein corona formation, protein corona isolation and mass spectrometry-based characterization. Each step presents unique challenges and interdependencies; flaws and a lack of essential process/protocol information in initial steps can lead to erroneous outcomes in proteomics characterization. This Perspective also highlights the importance of standardized protocols, discusses the advantages and shortcomings of current methodologies and proposes strategies to mitigate issues such as protein corona impurities and data reproducibility. By addressing these challenges, both academic and industrial nanomedicine and proteomics communities can enhance the reliability and accuracy of protein corona studies, thereby advancing the fields of nanomedicine and proteomics.</p>