Wang Junjie, Giordani Stefano, Marassi Valentina, Placci Anna, Roda Barbara, Reschiglian Pierluigi, Zattoni Andrea
Department of Chemistry G. Ciamician, University of Bologna, Bologna, 40126, Italy.
byFlow srl, Bologna, 40129, Italy.
Sci Rep. 2024 Dec 30;14(1):31568. doi: 10.1038/s41598-024-73624-0.
Gold nanoparticles (AuNPs) and their biocompatible conjugates find wide use as transducers in (bio)sensors and as Nano-pharmaceutics. The study of the interaction between AuNPs and proteins in representative application media helps to better understand their intrinsic behaviors. A multi-environment, multi-parameter screening strategy is proposed based on asymmetric flow field flow fractionation (AF4)-multidetector. Citrate-coated AuNPs ( AuCIT, 25.1 ± 0.2 nm) and PEG-coated AuNPs (AuPEG, 38.3 ± 0.8 nm) were employed with albumin as a model system. Attention was put in investigating the influence of Au/BSA mass ratios, that allowed to identify the yield-maximizing (1:1) and product-maximizing (2.5:1) conditions for the generation of AuNPs-protein conjugates. Furthermore, bioconjugate properties were thoroughly assessed across various saline media with different pH and ionic strengths. While AuNPs with PEG coating exhibit greater stability at high salinities, such as 30 mM, their conjugates are less stable over time. In contrast, although bare AuNPs are significantly affected by pH and salt concentration, once conjugates are formed, their stability surpasses that of the conjugates formed with AuPEG. The developed methodology can fill the vacancy of standard reference quality control (QC) procedures for bioconjugate synthesis and application in (bio)sensors and Nano-pharmaceutics, screening in a short time many combinations, easily scaling up to the semi-preparative scale or translating to different bioconjugates.
金纳米颗粒（AuNPs）及其生物相容性缀合物在（生物）传感器中作为换能器以及作为纳米药物有着广泛应用。研究AuNPs与代表性应用介质中蛋白质之间的相互作用有助于更好地理解它们的内在行为。基于不对称流场流分馏（AF4）-多检测器提出了一种多环境、多参数筛选策略。以柠檬酸盐包覆的AuNPs（AuCIT，25.1±0.2纳米）和聚乙二醇包覆的AuNPs（AuPEG，38.3±0.8纳米）与白蛋白作为模型系统。重点研究了Au/牛血清白蛋白质量比的影响，从而确定生成AuNPs-蛋白质缀合物的产量最大化（1:1）和产物最大化（2.5:1）条件。此外，还在不同pH值和离子强度的各种盐溶液介质中对生物缀合物的性质进行了全面评估。虽然聚乙二醇包覆的AuNPs在高盐浓度（如30 mM）下表现出更高的稳定性，但其缀合物随时间的稳定性较差。相比之下，尽管裸AuNPs受pH值和盐浓度的影响显著，但一旦形成缀合物，其稳定性超过了用AuPEG形成的缀合物。所开发的方法可以填补生物缀合物合成以及在（生物）传感器和纳米药物应用中的标准参考质量控制（QC）程序的空白，能在短时间内筛选多种组合，易于扩大到半制备规模或转化用于不同的生物缀合物。
