Protein Corona Stability and Removal from PET Microplastics: Analytical and Spectroscopic Evaluation in Simulated Intestinal Conditions

Microplastics entering the gastrointestinal environment rapidly acquire protein coronas that alter their surface chemistry and analytical detectability. We investigated the physicochemical interactions between fluorescently labeled bovine serum albumin (BSA) and polyethylene terephthalate (PET) microplastics during simulated intestinal exposure and evaluated the stability of the resulting hard corona. Using fluorescence tracking, SDS-PAGE, and FTIR spectroscopy, we showed that BSA forms a persistent corona that resists oxidative-only treatments. Only a combination of oxidation with an alkaline (KOH) or surfactant step (SDS) effectively removed the corona. None of the protocols applied affected polymer integrity. Residual protein in less effective protocols did not show changes on PET spectra in ATR FTIR. To validate the protocol under physiologically relevant complexity, we extended it to PET incubated with single digestive enzymes. FTIR spectra confirmed the removal of protein-specific signals in both systems, with no degradation of PET ester or aromatic functional groups nor signals of protein-polymer interactions. Our results highlight the robustness of protein-PET interactions in biological conditions and provide a variety of protocols for protein corona removal, suitable for diverse applications of microplastic analysis and toxicological studies.