Probing Nanoparticle-Protein Interaction by Capillary Electrophoresis
Probing Nanoparticle-Protein Interaction by Capillary Electrophoresis
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DOI：
10.1021/ac101627p
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发表时间：
2010-09-01
影响因子：
7.4
通讯作者：
Zhong, Wenwan
中科院分区：
文献类型：
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作者：
Li, Ni;Zeng, Shang;Zhong, Wenwan
了解纳米颗粒与蛋白质的相互作用有助于促进纳米颗粒在生物医学领域的应用，减少/预防纳米颗粒对生物系统可能造成的不良影响。纳米颗粒蛋白质相互作用的生物物理参数的定量测量将提高这样的理解，这可以方便地进行毛细管电泳（CE）在本研究中所示。确定了两种相互作用情况。通过毛细管区带电泳（CZE）从游离纳米颗粒和蛋白质中分离稳定的纳米颗粒-蛋白质复合物。具有快速缔合/解离速率的瞬时复合物在亲和毛细管电泳（ACE）中显示出与游离纳米颗粒不同的迁移率变化。牛血清白蛋白（BSA）与Fe 3 O 4纳米粒子（平均直径为8和10 nm）和与Au纳米粒子（平均直径为5和10 nm）的相互作用分别表现出缓慢和快速的结合动力学。利用Hill方程，计算了配合物的离解常数（K-D）和协同系数（n）。基于K-D和n值评价纳米颗粒和孵育缓冲液的理化性质对纳米颗粒蛋白相互作用的影响，以解释相互作用驱动力。我们的研究证明了CE在探测蛋白质与不同颗粒相互作用方面的高度简单性和灵活性。毛细管电泳的分离能力也应该促进多组分相互作用系统的研究，以研究吸附到纳米颗粒上如何影响蛋白质-蛋白质或蛋白质小分子相互作用。
Understanding nanoparticle-protein interaction could help in promoting applications of nanoparticles in the biomedical fields and reducing/preventing possible adverse effects to the biological systems caused by nanoparticles. Quantitative measurement of the biophysical parameters of nanoparticle protein interaction will improve such understanding, which could be conveniently performed by capillary electrophoresis (CE) as demonstrated in the present study. Two interaction situations were identified. Stable nanoparticle-protein complexes were resolved from the free nanoparticles and the proteins by capillary zone electrophoresis (CZE). Transient complexes with fast association/dissociation rates showed distinct mobility change from the free nanoparticles in affinity capillary electrophoresis (ACE). Interactions of bovine serum albumin (BSA) with the Fe3O4 nanoparticles (average diameters of 8 and 10 nm) and with the Au nanoparticles (average diameters of 5 and 10 nm) displayed slow and fast binding kinetics, respectively. Using the Hill equation, we could calculate the dissociation constants (K-D) and cooperativity coefficients (n). Impacts on nanoparticle protein interaction from the physicochemical properties of nanoparticles and the incubation buffer were evaluated on the basis of the K-D and n values to interpret the interaction driving forces. Our study demonstrated the high simplicity and flexibility of CE in probing the interaction of proteins with diverse particles. The separation power of CE should also facilitate studies of the multicomponent interaction systems for investigating how adsorption onto nanoparticles could affect the protein-protein or protein small molecule interactions.