Unidad de Investigación y Desarrollo, Probiomed, S.A. de C.V, Cruce de carreteras Acatzingo-Zumpahuacán S/N, C.P. 52400, Tenancingo, Estado de México, México.
Departamento de Ingeniería en Nanotecnología, Universidad de La Ciénega del Estado de Michoacán de Ocampo, Avenida Universidad 3000, C.P. 59103, Sahuayo, Michoacán, México.
AAPS PharmSciTech. 2018 May;19(4):1672-1680. doi: 10.1208/s12249-018-0983-6. Epub 2018 Mar 8.
Silver nanoparticles (AgNPs) are known to interact with proteins, leading to modifications of the plasmonic absorption that can be used to monitor this interaction, entailing a promising application for sensing adsorption of therapeutic proteins in primary containers. First, transmission electron microscopy in combination with plasmonic absorption and light scattering responses were used to characterize AgNPs and protein-AgNP complexes, including its concentration dependence, using two therapeutic molecules as models: a monoclonal antibody (mAb) and a synthetic copolymer (SC). Upon interaction, a protein corona was formed around AgNPs with the consequent shifting and broadening of their characteristic surface plasmon resonance (SPR) band (400 nm) to 410 nm and longer wavelenghts. Additional studies revealed secondary and three-dimensional structure modifications of model proteins upon interaction with AgNPs by circular dichroism and fluorescence techniques, respectively. Based on the modification of the SPR condition of AgNPs upon interaction with proteins, we developed a novel protein-sensing application of AgNPs in primary containers. This strategy was used to conduct a compatibility assessment of model proteins towards five commercially available prefillable glass syringe (PFS) models. mAb- and SC-exposed PFSs showed that 74 and 94% of cases were positive for protein adsorption, respectively. Interestingly, protein adsorption on 15% of total tested PFSs was negligible (below the nanogram level). Our results highlight the need of a case-by-case compatibility assessment of therapeutic proteins and their primary containers. This strategy has the potential to be easily applied on other containers and implemented during early-stage product development by pharmaceutical companies and for routine use during batch release by packaging manufacturers.
纳米银颗粒（AgNPs）已知会与蛋白质相互作用，导致等离子体吸收的修饰，可用于监测这种相互作用，为在初级容器中检测治疗性蛋白质的吸附提供了有前途的应用。首先，使用传输电子显微镜结合等离子体吸收和光散射响应来表征 AgNPs 和蛋白-AgNP 复合物，包括其浓度依赖性，使用两种治疗性分子作为模型：单克隆抗体（mAb）和合成共聚物（SC）。相互作用后，AgNPs 周围形成了蛋白质冠，导致其特征表面等离子体共振（SPR）带（400nm）移至 410nm 及更长波长，并变宽。进一步的研究通过圆二色性和荧光技术分别揭示了模型蛋白与 AgNPs 相互作用后的二级和三维结构修饰。基于 AgNPs 与蛋白质相互作用时 SPR 条件的变化，我们开发了一种 AgNPs 在初级容器中的新型蛋白质传感应用。该策略用于对五种市售预充式玻璃注射器（PFS）模型中的模型蛋白进行相容性评估。mAb 和 SC 暴露的 PFS 表明，分别有 74%和 94%的情况对蛋白质吸附呈阳性。有趣的是，在总测试的 PFS 中，有 15%的蛋白质吸附可以忽略不计（低于纳克水平）。我们的结果强调了需要对治疗性蛋白质及其初级容器进行逐个案例的相容性评估。这种策略有可能很容易地应用于其他容器，并在制药公司的早期产品开发阶段和包装制造商的常规批次放行期间实施。
