Department of Bioengineering, University of Illinois, Urbana-Champaign, Urbana, Illinois 61801, United States.
Biomedical Research Center, Carle Foundation Hospital, Urbana, Illinois 61801, United States.
ACS Appl Mater Interfaces. 2021 Oct 6;13(39):46464-46477. doi: 10.1021/acsami.1c17302. Epub 2021 Sep 27.
Many works utilize products isolated from nature as capping agents to functionalize gold nanoparticles for targeting and therapeutic applications. Some of the most advanced of these strategies utilize complex multicomponent biomaterials, such as whole cell-membranes, for nanoparticle functionalization strategies for evading or initializing immune response as well as for targeting. Strategies like these, wherein whole cell membrane is utilized for functionalization, take advantage of the complexity of the protein-lipid content and organization, which cells normally use for communication and interaction (instilling these capacities to nanoparticle vectors). Many approaches for achieving this in functionalizing the surface of nanoparticles rely on multistep processes, which necessitate the addition and then removal of synthetic molecules, heating, or pH modifications. These processes can have deleterious modifying effects on the functionalizing biomolecules, resulting in loss of product and time during each purification step, as well as potentially changing the biomolecule functionality toward a nondesirable effect. Here, we describe methods for forming gold nanoparticles at room temperature in a single step, functionalized with proteins, using nicotinamide adenine dinucleotide (NADH). This process enables formation of nanoparticles that can be functionalized by individual proteins (demonstrated with FBS) or whole cells membrane (extracted from B16F10 cells). This work is derivative from observations found in the literature by us and others, that mammalian cells are capable of producing gold nanoparticles from ionic gold without the supplementation of chemical species. The products of this single-step synthesis described herein have been optimized to maintain biomolecule integrity and so that there are no further purification steps required. To characterize the nanoparticles in terms of their shape, size, surface functionality, and biomolecule integrity throughout development, we employed light-based spectroscopy techniques, molecular modeling, electron microscopy, light scattering, and gel electrophoresis techniques. In order to compare the optimized biomolecule-functionalized nanoparticles against current standards (which require synthetic linkers, heating, or pH manipulation), we employed metabolic and live/dead assays as well as light-based microscopy/spectroscopy . In comparing our synthetic process against others for forming gold nanoparticles functionalized with complex biomolecule components (whole-cell membrane), we found that this process had superior particle internalization. Our strategy has similar outlets for application to these other works, however, because this process is entirely reliant on endogenous biomaterials and has additional potential.
许多工作利用从自然界中分离出来的产物作为帽状试剂，来修饰金纳米粒子，以用于靶向和治疗应用。其中一些最先进的策略利用复杂的多组分生物材料，如完整的细胞膜，来进行纳米粒子的功能化策略，以逃避或启动免疫反应，并进行靶向。像这样利用整个细胞膜进行功能化的策略，利用了蛋白质-脂质含量和组织的复杂性，细胞通常利用这些来进行通讯和相互作用（将这些能力注入纳米粒子载体）。许多实现这一目标的方法都依赖于多步过程，这些过程需要添加和去除合成分子、加热或 pH 修饰。这些过程可能会对功能化生物分子产生有害的修饰作用，导致每个纯化步骤中产品和时间的损失，并且可能会改变生物分子的功能，使其产生不理想的效果。在这里，我们描述了一种在室温下一步形成金纳米粒子的方法，该方法使用烟酰胺腺嘌呤二核苷酸（NADH）对其进行蛋白质功能化。该过程能够形成可以被单个蛋白质（用 FBS 进行演示）或整个细胞膜（从 B16F10 细胞中提取）功能化的纳米粒子。这项工作是从我们和其他人在文献中观察到的结果衍生而来的，哺乳动物细胞能够在没有添加化学物质的情况下从离子金中产生金纳米粒子。本文描述的单步合成的产物已经过优化，以保持生物分子的完整性，因此不需要进一步的纯化步骤。为了在整个开发过程中根据形状、尺寸、表面功能和生物分子完整性来表征纳米粒子，我们采用了基于光的光谱技术、分子建模、电子显微镜、光散射和凝胶电泳技术。为了将优化的生物分子功能化纳米粒子与当前的标准（需要合成接头、加热或 pH 处理）进行比较，我们采用了代谢和活/死测定以及基于光的显微镜/光谱学。在将我们的合成工艺与其他用于形成功能化金纳米粒子的复杂生物分子成分（完整细胞膜）的工艺进行比较时，我们发现该工艺具有优越的颗粒内化能力。我们的策略在应用于这些其他工作方面具有相似的出口，但由于该工艺完全依赖于内源性生物材料，因此具有额外的潜力。
