Kurup Chitra P, Mohd-Naim Noor F, Keasberry Natasha A, Zakaria Siti N A, Bansal Vipul, Ahmed Minhaz U
Biosensors and Nanobiotechnology Laboratory, Integrated Science Building, Faculty of Science, Universiti Brunei Darussalam, Jalan Tungku Link, GadongBE 1410, Brunei Darussalam.
PAPRSB Institute of Health Sciences, Universiti Brunei Darussalam, Jalan Tungku Link, GadongBE 1410, Brunei Darussalam.
ACS Omega. 2022 Oct 20;7(43):38709-38716. doi: 10.1021/acsomega.2c04300. eCollection 2022 Nov 1.
A molybdenum sulfide/zirconium oxide/Nafion (MoS/ZrO/Naf) based electrochemiluminescence (ECL) aptasensor for the selective and ultrasensitive detection of ApoA1 is proposed, with Ru(bpy) as the luminophore. The chitosan (CS) modification on the nanocomposite layer allowed glutaraldehyde (GLUT) cross-linking, resulting in the immobilization of ApoA1 aptamers. Scanning electron microscopy, tunneling electron microscopy, and energy dispersive X-ray spectroscopy were used to characterize the nanocomposite, while electrochemiluminescence (ECL), cyclic voltammetry, and electrochemical impedance spectroscopy were used to analyze the aptasensor assembly. The nanocomposite was used as an electrode modifier, which increased the intensity of the ECL signal. Due to the anionic environment produced on the sensor surface following the specific interaction of the ApoA1 biomarker with the sensor, more Ru(bpy) were able to be electrostatically attached to the aptamer-ApoA1 complex, resulting in enhanced ECL signal. The ECL aptasensor demonstrated outstanding sensitivity for ApoA1 under optimal experimental conditions, with a detection limit of 53 fg/mL and a wide linear dynamic range of 0.1-1000 pg/mL. The potential practical applicability of this aptasensor was validated by analyzing ApoA1 in human serum samples, with recovery rates of 94-108% ( = 3). The proposed assay was found to be substantially better compared to the commercially available enzyme-linked immunosorbent assay method, as reflected from over 1500 times improvement in the detection limit for ApoA1.
提出了一种基于硫化钼/氧化锆/全氟磺酸(MoS/ZrO/Naf)的电化学发光(ECL)适体传感器,用于选择性和超灵敏检测载脂蛋白A1(ApoA1),以钌(联吡啶)[Ru(bpy)]作为发光体。纳米复合层上的壳聚糖(CS)修饰允许戊二醛(GLUT)交联,从而实现ApoA1适体的固定。使用扫描电子显微镜、隧道电子显微镜和能量色散X射线光谱对纳米复合材料进行表征,同时使用电化学发光(ECL)、循环伏安法和电化学阻抗谱对适体传感器组件进行分析。该纳米复合材料用作电极修饰剂,提高了ECL信号强度。由于ApoA1生物标志物与传感器特异性相互作用后在传感器表面产生阴离子环境,更多的Ru(bpy)能够静电附着在适体 - ApoA1复合物上,导致ECL信号增强。该ECL适体传感器在最佳实验条件下对ApoA1表现出出色的灵敏度,检测限为53 fg/mL,线性动态范围宽达0.1 - 1000 pg/mL。通过分析人血清样本中的ApoA1验证了该适体传感器的潜在实际适用性,回收率为94 - 108%(n = 3)。与市售酶联免疫吸附测定方法相比,该方法有显著优势,ApoA1检测限提高了1500倍以上。