Direct Quantification of Serum Protein Interactions with PEGylated Micelle Nanocarriers
Direct Quantification of Serum Protein Interactions with PEGylated Micelle Nanocarriers
复制标题
血清蛋白与聚乙二醇化胶束纳米载体相互作用的直接定量
DOI：
10.1021/acs.biomac.2c01538
复制
发表时间：
2023
影响因子：
6.2
通讯作者：
Smith, Adam W.
中科院分区：
文献类型：
--
作者：
Mallory, D. Paul;Freedman, Abegel;Kaliszewski, Megan J.;Montenegro-Galindo, Gladys Rocío;Pugh, Coleen;Smith, Adam W.
存在大量的纳米载体(NC)技术，每一种技术在靶向性、稳定性和免疫学惰性方面都具有高度特定的优势。在生理条件下表征这些纳米特性对于开发优化的药物输送系统是至关重要的。一种通过避免蛋白质吸附在NCS上来减少过早消除的方法是用聚乙二醇(PEG)进行表面功能化，恰当地称为聚乙二醇化。然而，最近的研究表明，一些聚乙二醇化的NC具有延迟的免疫反应，表明存在蛋白质与NC的相互作用。明显的蛋白质-NC相互作用，特别是在胶束体系中，可能被忽视了，因为许多早期的研究依赖于对分子水平相互作用不那么敏感的技术。已经开发了更灵敏的技术，但主要的挑战是直接测量相互作用，这必须在现场进行，因为胶束组装是动态的。在这里，我们报告了使用脉冲交错激发荧光互相关光谱(PIE-FCCS)来询问两个基于聚乙二醇基的胶束模型与血清白蛋白之间的相互作用，以比较基于线性或循环聚乙二醇组分的蛋白质吸附差异。首先，通过测量胶束在孤立和混合溶液中的扩散，我们证实了两嵌段和三嵌段共聚物胶束组装的热稳定性。此外，我们测量了胶束和血清蛋白的共扩散，其大小随着浓度和继续孵育的增加而增加。结果表明，PIE-FCCS能够测量荧光标记的NC与血清蛋白之间的直接相互作用，即使在比生理观察浓度低500倍的情况下也是如此。这一能力展示了PIE-FCC在仿生条件下表征药物输送系统的潜在效用。
A large repertoire of nanocarrier (NC) technologies exists, each with highly specified advantages in terms of targetability, stability, and immunological inertness. The characterization of such NC properties within physiological conditions is essential for the development of optimized drug delivery systems. One method that is well established for reducing premature elimination by avoiding protein adsorption on NCs is surface functionalization with poly(ethylene glycol) (PEG), aptly called PEGylation. However, recent studies revealed that some PEGylated NCs have a delayed immune response, indicating the occurrence of protein-NC interactions. Obvious protein-NC interactions, especially in micellar systems, may have been overlooked as many early studies relied on techniques less sensitive to molecular level interactions. More sensitive techniques have been developed, but a major challenge is the direct measurement of interactions, which must be done in situ, as micelle assemblies are dynamic. Here, we report the use of pulsed-interleaved excitation fluorescence cross-correlation spectroscopy (PIE-FCCS) to interrogate the interactions between two PEG-based micelle models and serum albumin protein to compare protein adsorption differences based on linear or cyclic PEG architectures. First, by measuring micelle diffusion in isolated and mixed solutions, we confirmed the thermal stability of diblock and triblock copolymer micelle assemblies. Further, we measured the co-diffusion of micelles and serum proteins, the magnitudes of which increased with concentration and continued incubation. The results demonstrate that PIE-FCCS is capable of measuring direct interactions between fluorescently labeled NC and serum proteins, even at concentrations 500 times lower than those observed physiologically. This capability showcases the potential utility of PIE-FCCS in the characterization of drug delivery systems in biomimetic conditions.