Abstract：
Improving the clinical translation of nanomedicine requires better knowledge about how nanoparticles interact with biological environments. As researchers are recognizing the importance of understanding the protein corona and characterizing how nanocarriers respond in biological systems, new tools and techniques are needed to analyze nanocarrier-protein interactions, especially for smaller size (<10 nm) nanoparticles like polyamidoamine (PAMAM) dendrimers. Here, we developed a streamlined, semiquantitative approach to assess dendrimer-protein interactions using a nondenaturing electrophoresis technique combined with mass spectrometry. With this protocol, we detect fluorescently tagged dendrimers and proteins simultaneously, enabling us to analyze when dendrimers migrate with proteins. We found that PAMAM dendrimers mostly interact with complement proteins, particularly C3 and C4a, which aligns with previously published data, verifying that our approach can be used to isolate and identify dendrimer-protein interactions.
摘要：
改善纳米医学的临床翻译需要更好地了解纳米颗粒如何与生物环境相互作用。随着研究人员认识到理解蛋白质电晕和表征纳米载体在生物系统中反应的重要性，需要新的工具和技术来分析纳米载体-蛋白质相互作用，特别是对于较小尺寸(<10nm)的纳米颗粒,如聚酰胺胺(PAMAM)树枝状聚合物。这里,我们开发了一个流线型的，半定量方法使用非变性电泳技术结合质谱来评估树枝状聚合物-蛋白质相互作用。有了这个协议，我们同时检测荧光标记的树枝状聚合物和蛋白质，使我们能够分析树枝状聚合物何时随蛋白质迁移。我们发现PAMAM树枝状聚合物主要与补体蛋白相互作用，特别是C3和C4a，与以前发布的数据一致，验证我们的方法可用于分离和鉴定树枝状聚合物-蛋白质相互作用。
