PEG modification enhances the in vivo stability of bioactive proteins immobilized on magnetic nanoparticles

Objective To increase the in vivo stability of bioactive proteins via optimized loading methods. Results β-Glucosidase (β-Glu), as a model protein, was immobilized on magnetic nanoparticles(denoted as MNP-β-Glu) by chemical coupling methods and was further modified by poly(ethylene glycol) (PEG) molecules (denoted as MNP-β-Glu-PEG) to increase its stability. The physicochemical properties of the as-prepared nanohybrids, including the particle size, zeta potential, and enzyme activity, were well characterized. The proper MNP/β-Glu feed ratio was important for optimizing the particle size. Analysis of enzyme activity showed that the stability of immobilized β-Glu compared with free β-Glu was lower in deionized water and higher in blood serum at 37 °C. MNP-β-Glu-PEG retained 77.9% of the initial activity within 30 days at 4 °C, whereas the free enzyme retained only 58.2%. Pharmacokinetic studies of Sprague–Dawley (SD) rats showed that the MNP-β-Glu-PEG group retained a higher enzyme activity in vivo (41.46% after 50 min) than the MNP-β-Glu group (0.03% after 50 min) and the β-Glu group (0.37% after 50 min). Moreover, in contrast to the MNP-β-Glu group, the enzyme activity was not fully synchronous with the decrease in the Fe concentration in the MNP-β-Glu-PEG group. Conclusions All findings indicated that the method of immobilization on magnetic nanoparticles and PEG modification is promising for the application of bioactive proteins in vivo.