The Key Laboratory of Functional Molecular Solids, Ministry of Education; Anhui Province Key Laboratory of Biomedical Materials and Chemical Measurement; College of Chemistry and Materials Science, Anhui Normal University, Wuhu, 241002, P. R. China.
Mol Pharm. 2024 Nov 4;21(11):5598-5606. doi: 10.1021/acs.molpharmaceut.4c00552. Epub 2024 Oct 24.
The accurate and efficient quantification of nanodrug dosage is crucial for early anticancer therapy. The enzyme-linked immunosorbent assay (ELISA) has emerged as a robust tool for detecting anticancer nanodrug dosage; however, the development of sensing elements to quantify anticancer nanodrugs still poses a challenge. To overcome this problem, we utilize polysuccinimide-loaded curcumin (CUR @PSI) as a model to employ an ELISA based on peroxidase nanozyme Pt-SiO Janus nanoparticles (Pt-SiO JNPs) for the indirect quantitative analysis of intracellular anticancer nanodrug dosage. This novel approach employs an immunoassay to indirectly quantify the dosage of anticancer nanodrugs while preserving its structural integrity. The silica components of Pt-SiO JNPs adsorb intermediates, while the Pt NP components exhibit high catalytic activity. Pt-SiO JNPs are functionalized with anti-PSI antibody (Pt-SiO JNPs-Ab) to serve as an immunosensor capable of specific recognition of CUR @PSI. Additionally, we employed cytotoxicity assays and confocal imaging techniques to demonstrate the excellent biocompatibility of CUR @PSI, as well as its specific uptake by cancer cells. According to the experimental results, the limit of detection (LOD) for the immunoassay of Pt-SiO JNPs as a marker for detecting CUR @PSI is approximately 4.5-fold lower than that of horseradish peroxidase. Therefore, by optimizing the conditions, we established a direct competitive ELISA using Pt-SiO JNPs as colorimetric indicators for the quantitative detection of intracellular CUR @PSI. The LOD for this ELISA was determined to be 0.01 ng/mL, while the loaded CUR amount calculated from the drug loading capacity was found to be 0.22 pg/mL. Furthermore, the recoveries obtained from this established ELISA ranged between 94.0 and 108%, demonstrating excellent accuracy. Consequently, the peroxidase mimic Pt-SiO JNPs-based ELISA exhibits significant potential for precise quantification of intracellular anticancer nanodrug dosages.
准确高效地定量纳米药物剂量对于早期抗癌治疗至关重要。酶联免疫吸附测定（ELISA）已成为检测抗癌纳米药物剂量的强大工具；然而，开发用于定量抗癌纳米药物的传感元件仍然是一个挑战。为了克服这个问题，我们利用负载姜黄素的聚琥珀酰亚胺（CUR@PSI）作为模型，采用基于过氧化物酶纳米酶 Pt-SiO 二聚体纳米粒子（Pt-SiO JNPs）的 ELISA 间接定量分析细胞内抗癌纳米药物剂量。这种新方法采用免疫测定法间接定量抗癌纳米药物的剂量，同时保持其结构完整性。Pt-SiO JNPs 的二氧化硅部分吸附中间体，而 Pt NP 部分则表现出高催化活性。Pt-SiO JNPs 被抗 PSI 抗体（Pt-SiO JNPs-Ab）功能化，作为能够特异性识别 CUR@PSI 的免疫传感器。此外，我们还采用细胞毒性测定和共聚焦成像技术证明了 CUR@PSI 的优异生物相容性及其被癌细胞的特异性摄取。根据实验结果，作为检测 CUR@PSI 的标记物，Pt-SiO JNPs 的免疫测定的检测限（LOD）比辣根过氧化物酶低约 4.5 倍。因此，通过优化条件，我们建立了一种直接竞争 ELISA，使用 Pt-SiO JNPs 作为比色指示剂用于定量检测细胞内 CUR@PSI。该 ELISA 的 LOD 确定为 0.01 ng/mL，而从药物载药量计算得出的负载 CUR 量为 0.22 pg/mL。此外，该建立的 ELISA 的回收率在 94.0%至 108%之间，表现出优异的准确性。因此，基于过氧化物酶模拟物 Pt-SiO JNPs 的 ELISA 具有精确定量细胞内抗癌纳米药物剂量的巨大潜力。
