BioMark/ISEP, School of Engineering of the Polytechnic Institute of Porto, Porto, Portugal.
LAQV/REQUIMTE, Faculty of Pharmacy of Porto University, Porto, Portugal.
Sci Rep. 2018 Mar 21;8(1):4944. doi: 10.1038/s41598-018-23271-z.
A highly sensitive fluorescence detection probe was developed by tailoring plastic antibodies on the external surface of aqueous soluble quantum dots (QDs). The target was Myoglobin (Myo), a cardiac biomarker that quenched the intrinsic fluorescent emission of cadmium telluride (CdTe) QDs capped with mercaptopropionic acid (CdTe-MPA-QDs). The QDs were incubated with the target protein and further modified with a molecularly-imprinted polymer (MIP) produced by radical polymerization of acrylamide and bisacrylamide. The main physical features of the materials were assessed by electron microscopy, dynamic light scattering (DLS), UV/Vis spectrophotometry and spectrofluorimetry. The plastic antibodies enabled Myo rebinding into the QDs with subsequent fluorescence quenching. This QD-probe could detect Myo concentrations from 0.304 to 571 pg/ml (50.6 fM to 95 pM), with a limit of detection of 0.045 pg/ml (7.6 fM). The proposed method was applied to the determination of Myo concentrations in synthetic human serum. The results obtained demonstrated the ability of the modified-QDs to determine Myo below the cut-off values of myocardial infarction. Overall, the nanostructured MIP-QDs reported herein displayed quick responses, good stability and sensitivity, and high selectivity for Myo, offering the potential to be explored as new emerging sensors for protein detection in human samples.
通过在水溶性量子点(QDs)的外表面定制塑料抗体,开发了一种高灵敏度的荧光检测探针。该探针的目标是肌红蛋白(Myo),一种心脏生物标志物,可猝灭巯基丙酸(CdTe-MPA-QDs)封端的碲化镉(CdTe)QDs 的本征荧光发射。将 QDs 与靶蛋白孵育,并用丙烯酰胺和双丙烯酰胺的自由基聚合产生的分子印迹聚合物(MIP)进一步修饰。通过电子显微镜、动态光散射(DLS)、紫外/可见分光光度法和荧光分光光度法评估了材料的主要物理特性。塑料抗体使 Myo 重新结合到 QDs 中,随后发生荧光猝灭。该 QD 探针可检测 0.304 至 571 pg/ml(50.6 fM 至 95 pM)的 Myo 浓度,检测限为 0.045 pg/ml(7.6 fM)。该方法已应用于合成人血清中 Myo 浓度的测定。结果表明,修饰后的 QDs 能够在心肌梗死的截止值以下测定 Myo。总的来说,本文报道的纳米结构 MIP-QDs 具有快速响应、良好的稳定性和灵敏度以及对 Myo 的高选择性,有望作为用于检测人样品中蛋白质的新型新兴传感器进行探索。