Concentration and composition of the protein corona as a function of incubation time and serum concentration: an automated approach to the protein corona
Concentration and composition of the protein corona as a function of incubation time and serum concentration: an automated approach to the protein corona
复制标题
DOI：
10.1007/s00216-022-04278-y
复制
发表时间：
2022-08-26
影响因子：
4.3
通讯作者：
Payne, Christine K.
中科院分区：
文献类型：
--
作者：
Poulsen, Karsten M.;Payne, Christine K.
与蛋白质接触的纳米颗粒形成吸附在纳米颗粒表面的蛋白质“冠”。随后的生物反应是由吸附的蛋白质而不是裸露的纳米颗粒介导的。如果研究人员能够预测哪些特定蛋白质将吸附在纳米颗粒表面上，纳米颗粒作为纳米药物和生物传感器的使用将得到极大改善。我们使用最近开发的带有液体处理机器人和低成本蛋白质组学的自动化工作流程来确定羧酸盐修饰的氧化铁纳米粒子（200 nm）上形成的蛋白质电晕的浓度和组成，作为孵育时间和血清浓度的函数。我们通过比色测定法测量所得蛋白冠的浓度，并通过蛋白质组学测量蛋白冠的组成，报告相对于用于形成冠的胎牛血清 (FBS) 蛋白的丰度和富集度。研究发现，在高（100% FBS）孵育浓度下，孵育时间是电晕浓度和组成的一个重要参数，而在低（10%）FBS 浓度下仅产生轻微影响。除了这些发现之外，我们还描述了两种方法学进展，以帮助降低与蛋白质电晕实验相关的成本。我们已经自动化了蛋白质组学所需的消化步骤，并测量了蛋白质组学实验每个阶段的一式三份样品之间的变异性。总的来说，这些结果证明了了解影响电晕形成的多个参数、为电晕表征提供新工具以及推进纳米材料生物分析研究的重要性。
Nanoparticles in contact with proteins form a "corona" of proteins adsorbed on the nanoparticle surface. Subsequent biological responses are then mediated by the adsorbed proteins rather than the bare nanoparticles. The use of nanoparticles as nanomedicines and biosensors would be greatly improved if researchers were able to predict which specific proteins will adsorb on a nanoparticle surface. We use a recently developed automated workflow with a liquid handling robot and low-cost proteomics to determine the concentration and composition of the protein corona formed on carboxylate-modified iron oxide nanoparticles (200 nm) as a function of incubation time and serum concentration. We measure the concentration of the resulting protein corona with a colorimetric assay and the composition of the corona with proteomics, reporting both abundance and enrichment relative to the fetal bovine serum (FBS) proteins used to form the corona. Incubation time was found to be an important parameter for corona concentration and composition at high (100% FBS) incubation concentrations, with only a slight effect at low (10%) FBS concentrations. In addition to these findings, we describe two methodological advances to help reduce the cost associated with protein corona experiments. We have automated the digest step necessary for proteomics and measured the variability between triplicate samples at each stage of the proteomics experiments. Overall, these results demonstrate the importance of understanding the multiple parameters that influence corona formation, provide new tools for corona characterization, and advance bioanalytical research in nanomaterials.