Formation kinetics, stability, and field-mediated interaction with biological systems of a protein corona generated on magnetic nanoparticles

磁性纳米粒子上生成的蛋白质电晕的形成动力学、稳定性以及与生物系统的场介导相互作用

• 批准号: 238058170
• 负责人: Professor Dr.-Ing. Silvio Dutz
• 金额: --
• 依托单位: Leupold-Institut für Angewandte Naturwissenschaften (LIAN)
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2013
• 资助国家: 德国
• 起止时间: 2012-12-31 至 2018-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/238058170?language=en
• 关键词: Formation; kinetics; stability; field; mediated

This project focuses on the controlled formation of a protein corona on magnetic nanoparticles (MNP), the influence of different factors onto this process, as well as the influence of the formed corona on interactions of these hybrid particles with a biological system. We will therefore synthesize core-shell hybrid particles, consisting of a magnetic core and a polymer shell of varying charge. These hybrid particles then are incubated in fetal calve serum (FSC) for the formation of a protein corona. We are particularly interested in elucidating the interaction of particles after incubation with different biological systems. This is investigated by monitoring the contact between MNP@corona hybrid particles with blood, different cell lines, and animals - in certain cases also under the influence of an external field or field gradient. Suitable methods are µ-rheology, established cell toxicity assays, as well as 2D and 3D microscopic techniques for cell lines and animals. Further, we are also interested in how such particles have to be prepared or stored for in-vivo applications. Another important goal of this project is to gain in-depth understanding about the kinetics of protein corona formation. For this, key technologies are magnetic relaxometry, AC-susceptometry, and small angle X-ray scattering (SAXS) - all of which are highly sensitive towards small changes in hydrodynamic diameter and, hence, the growth of the protein corona. During the first period of this priority program, we established techniques for a defined and reproducible corona formation. We are now interested in probing mechanical properties as well as the (chemical) stability of the protein corona. This is realized by incorporating two dyes representing a FRET pair (Förster resonance energy transfer) within polymer shell and protein corona. As long as the corona is intact, both counterparts are in close proximity and FRET can be detected. Upon release of the protein corona, this energy transfer disappears and we plan to use this for probing corona stability under different conditions, including in-vivo experiments. In another part of this project, we investigate the effect of a compartmentalized polymer shell, e.g. through the presence of hydrophobic and hydrophilic patches, onto the protein corona formation. We anticipate that using core-shell particles featuring such patches influences both the mechanism of protein adsorption as well as the amount and composition of the corona being formed.

该项目的重点是在磁性纳米颗粒（MNP）上控制蛋白质冠的形成，不同因素对这一过程的影响，以及形成的冠对这些混合颗粒与生物系统相互作用的影响。因此，我们将合成核壳混合粒子，由磁性核和不同电荷的聚合物壳组成。然后将这些杂合颗粒在胎牛血清（FSC）中孵育以形成蛋白质冠。我们特别感兴趣的是阐明粒子与不同生物系统孵育后的相互作用。这是通过监测MNP@冠状杂交颗粒与血液、不同细胞系和动物之间的接触来研究的-在某些情况下还在外场或场梯度的影响下。合适的方法是微流变学、已建立的细胞毒性测定以及用于细胞系和动物的2D和3D显微镜技术。此外，我们还对如何制备或储存这些颗粒以用于体内应用感兴趣。该项目的另一个重要目标是深入了解蛋白质冠形成的动力学。为此，关键技术是磁弛豫法，AC-散射法和小角X射线散射（SAXS）-所有这些都对流体动力学直径的微小变化高度敏感，因此，蛋白质冠的生长。在这个优先项目的第一阶段，我们建立了一个明确的和可重复的电晕形成技术。我们现在感兴趣的是探测蛋白质冠的机械特性以及（化学）稳定性。这是通过在聚合物壳和蛋白质冠内掺入代表FRET对（福斯特共振能量转移）的两种染料来实现的。只要冠是完整的，两个对应物是在密切的接近和FRET可以检测到。蛋白质冠释放后，这种能量转移消失，我们计划使用它来探测冠在不同条件下的稳定性，包括体内实验。在这个项目的另一部分，我们研究了一个区室化的聚合物壳的影响，例如通过疏水和亲水补丁的存在，对蛋白质冠形成。我们预计，使用具有这种补丁的核壳颗粒影响蛋白质吸附的机制以及形成的电晕的量和组成。