Radiology Department, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan, 430000, Hubei, China.
Hubei Province Academy of Traditional Chinese Medicine, Wuhan, 430000, Hubei, China.
Biotechnol Lett. 2020 Aug;42(8):1407-1418. doi: 10.1007/s10529-020-02867-4. Epub 2020 Mar 21.
To increase the in vivo stability of bioactive proteins via optimized loading methods.
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.
All findings indicated that the method of immobilization on magnetic nanoparticles and PEG modification is promising for the application of bioactive proteins in vivo.
通过优化载药方法提高生物活性蛋白的体内稳定性。
以β-葡萄糖苷酶（β-Glu）为模型蛋白，通过化学偶联法将其固定在磁性纳米颗粒上（记为 MNP-β-Glu），然后进一步用聚乙二醇（PEG）分子进行修饰（记为 MNP-β-Glu-PEG），以提高其稳定性。所制备的纳米杂化物的物理化学性质，包括粒径、Zeta 电位和酶活性，都得到了很好的表征。适当的 MNP/β-Glu 进料比对于优化粒径很重要。酶活性分析表明，与游离β-Glu 相比，固定化β-Glu 在去离子水和 37°C 血清中的稳定性较低。MNP-β-Glu-PEG 在 4°C 下 30 天内保留了 77.9%的初始活性，而游离酶仅保留了 58.2%。Sprague-Dawley（SD）大鼠的药代动力学研究表明，MNP-β-Glu-PEG 组在体内保持了更高的酶活性（50 分钟后为 41.46%），高于 MNP-β-Glu 组（50 分钟后为 0.03%）和β-Glu 组（50 分钟后为 0.37%）。此外，与 MNP-β-Glu 组相比，MNP-β-Glu-PEG 组的酶活性与 Fe 浓度的下降并不完全同步。
所有发现表明，磁性纳米颗粒固定化和 PEG 修饰的方法有望将生物活性蛋白应用于体内。
