Lu Minglei, He Qiyi, Zhong Yingying, Pan Junkang, Lao Zhiting, Lin Mingxia, Wang Tiantian, Cui Xiping, Ding Jinlong, Zhao Suqing
Department of Pharmaceutical Engineering, School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou 510006, People's Republic of China.
Anal Methods. 2021 Mar 11;13(9):1164-1171. doi: 10.1039/d1ay00053e.
Progesterone (P4) belongs to a factor that affects stress response and is a potential carcinogen, and saliva levels are expected to be a standard measurement for clinical diagnosis. In this study, a new type of nanoflower with both recognition functionality and catalytic substrate ability was prepared by copper phosphate, Pt/IrO2 nanocomposites (Pt/IrO2 NPs), streptavidin (SA) and horseradish peroxidase (HRP) via a one-pot co-precipitation strategy. Due to the enhanced catalytic activity and stability of Pt/IrO2@SA@HRP nanoflowers, we developed a powerful and sensitive multiple-catalysis ELISA to monitor progesterone in saliva. Multiple-catalysis ELISA based on a specific antibody and Pt/IrO2@SA@HRP nanoflowers exhibited a linear interval range from 0.217 ng mL-1 to 7.934 ng mL-1. The median inhibitory concentration (IC50) for progesterone is 1.311 ng mL-1 and the limit of detection (LOD = IC10) is 0.076 ng mL-1 in the proposed method. Satisfactory recoveries were in a range of 79.6-107% with an acceptable coefficient of variation (below 10.6%). Results of the multiple-catalysis ELISA and LC-MS/MS had a good coincidence. Our result unraveled that multiple-catalysis ELISA is a potentially serviceable tool for the detection of progesterone in saliva.
孕酮（P4）属于影响应激反应的一种因子，且是一种潜在致癌物，唾液水平有望成为临床诊断的标准检测指标。在本研究中，通过一锅共沉淀策略，利用磷酸铜、铂/二氧化铱纳米复合材料（Pt/IrO2 NPs）、链霉亲和素（SA）和辣根过氧化物酶（HRP）制备了一种兼具识别功能和催化底物能力的新型纳米花。由于Pt/IrO2@SA@HRP纳米花的催化活性和稳定性增强，我们开发了一种强大且灵敏的多重催化酶联免疫吸附测定法来监测唾液中的孕酮。基于特异性抗体和Pt/IrO2@SA@HRP纳米花的多重催化酶联免疫吸附测定法的线性区间范围为0.217 ng mL-1至7.934 ng mL-1。在所提出的方法中，孕酮的半数抑制浓度（IC50）为1.311 ng mL-1，检测限（LOD = IC10）为0.076 ng mL-1。回收率在79.6%至107%范围内，变异系数可接受（低于10.6%）。多重催化酶联免疫吸附测定法和液相色谱-串联质谱法的结果具有良好的一致性。我们的结果表明，多重催化酶联免疫吸附测定法是检测唾液中孕酮的一种潜在可用工具。
