Białobrzeska Wioleta, Firganek Daniel, Czerkies Maciej, Lipniacki Tomasz, Skwarecka Marta, Dziąbowska Karolina, Cebula Zofia, Malinowska Natalia, Bigus Daniel, Bięga Ewelina, Pyrć Krzysztof, Pala Katarzyna, Żołędowska Sabina, Nidzworski Dawid
Institute of Biotechnology and Molecular Medicine, 3 Trzy Lipy St., 80-172 Gdansk, Poland.
SensDx, 14b Postępu St., 02-676 Warszawa, Poland.
Biosensors (Basel). 2020 Nov 13;10(11):175. doi: 10.3390/bios10110175.
This paper presents the development and comparison of label-free electrochemical immunosensors based on screen-printed gold and glassy carbon (GC) disc electrodes for efficient and rapid detection of respiratory syncytial virus (RSV). Briefly, the antibody specific to the F protein of RSV was successfully immobilized on modified electrodes. Antibody coupling on the Au surface was conducted via 4-aminothiophenol (4-ATP) and glutaraldehyde (GA). The GC surface was modified with poly-L-lysine (PLL) for direct anti-RSV conjugation after EDC/NHS (1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide/N-Hydroxysuccinimide) activation. Electrochemical characterizations of the immunosensors were carried out by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). GC-based immunosensors show a dynamic range of antigen detection from 1.0 × 10 PFU/mL to 1.5×10 PFU/mL, more than 1.0 × 10 PFU/mL to 1.0 × 10 PFU/mL for the Au-based sensor. However, the GC platform is less sensitive and shows a higher detection limit (LOD) for RSV. The limit of detection of the Au immunosensor is 1.1 × 10 PFU/mL, three orders of magnitude lower than 2.85 × 10 PFU/mL for GC. Thus, the Au-based immunosensor has better analytical performance for virus detection than a carbon-based platform due to high sensitivity and very low RSV detection, obtained with good reproducibility.
本文介绍了基于丝网印刷金电极和玻碳(GC)圆盘电极的无标记电化学免疫传感器的开发与比较,用于高效快速检测呼吸道合胞病毒(RSV)。简而言之,成功地将针对RSV F蛋白的抗体固定在修饰电极上。通过4-氨基硫酚(4-ATP)和戊二醛(GA)在金表面进行抗体偶联。在经EDC/NHS(1-乙基-3-(3-二甲基氨基丙基)碳二亚胺/N-羟基琥珀酰亚胺)活化后,用聚-L-赖氨酸(PLL)修饰GC表面以直接进行抗RSV偶联。通过循环伏安法(CV)和电化学阻抗谱(EIS)对免疫传感器进行电化学表征。基于GC的免疫传感器显示抗原检测的动态范围为1.0×10 PFU/mL至1.5×10 PFU/mL,基于金的传感器则为1.0×10 PFU/mL至1.0×10 PFU/mL以上。然而,GC平台的灵敏度较低,对RSV的检测限(LOD)较高。金免疫传感器的检测限为1.1×10 PFU/mL,比GC的2.85×10 PFU/mL低三个数量级。因此,基于金的免疫传感器由于具有高灵敏度、极低的RSV检测限且重现性良好,在病毒检测方面比基于碳的平台具有更好的分析性能。