Adhikari Juthi, Keasberry Natasha Ann, Mahadi Abdul Hanif, Yoshikawa Hiroyuki, Tamiya Eiichi, Ahmed Minhaz Uddin
Biosensors and Biotechnology Laboratory, Chemical Science Programme, Faculty of Science, Universiti Brunei Darussalam Jalan Tungku Link, Gadong BE 1410 Brunei Darussalam
Centre for Advanced Material and Energy Sciences, Universiti Brunei Darussalam Tungku Link, Gadong BE1410 Brunei Darussalam.
RSC Adv. 2019 Oct 24;9(59):34283-34292. doi: 10.1039/c9ra05016g. eCollection 2019 Oct 23.
This study presents a novel and ultrasensitive electrochemiluminescence approach for the quantitative assessment of creatine kinase MB (CK-MB). Both carbon, carbon nano-onions (CNOs) and metal-based nanoparticles, such as gold nanoparticles (AuNPs) and iron oxide (FeO), were combined to generate a unique nanocomposite for the detection of CKMB. The immunosensor construction involved the deposition of the nanocomposite on the working electrode, followed by the incubation of an antibody and a blocking agent. Tris(2,2'-bipyridyl)-ruthenium(ii) chloride ([Ru(bpy)]Cl) was used as a luminophore, where tri--propylamine (TPrA) was selected as the co-reactant due to its aqueous immobility and luminescence properties. The analytical performance was demonstrated by cyclic voltammetry on ECL. The characterization of each absorbed layer was performed by cyclic voltammetry (CV) and chronocoulometry (CC) techniques in both EC and ECL. For further characterization of iron oxide, gold nanoparticles and carbon nano-onions, scanning electron microscopy (SEM), transmission electron microscopy (TEM) and X-ray diffraction (XRD) were performed. The proposed immunosensor showcases a wide linear range (10 ng mL to 50 fg mL), with an extremely low limit of detection (5 fg mL). This CKMB immunosensor also exhibits remarkable selectivity, reproducibility, stability and resistance capability towards common interferences available in human serum. In addition, the immunosensor holds great potential to work with real serum samples for clinical diagnosis.
本研究提出了一种用于定量评估肌酸激酶同工酶MB（CK-MB）的新型超灵敏电化学发光方法。碳、碳纳米洋葱（CNOs）以及金属基纳米颗粒，如金纳米颗粒（AuNPs）和氧化铁（FeO），被组合在一起以生成一种用于检测CKMB的独特纳米复合材料。免疫传感器的构建包括将纳米复合材料沉积在工作电极上，随后孵育抗体和封闭剂。三（2,2'-联吡啶）钌（II）氯化物（[Ru(bpy)]Cl）用作发光体，由于其在水中的不移动性和发光特性，选择三丙胺（TPrA）作为共反应剂。通过循环伏安法对电化学发光进行了分析性能验证。通过循环伏安法（CV）和计时库仑法（CC）技术在电化学和电化学发光中对每个吸附层进行了表征。为了进一步表征氧化铁、金纳米颗粒和碳纳米洋葱，进行了扫描电子显微镜（SEM）、透射电子显微镜（TEM）和X射线衍射（XRD）分析。所提出的免疫传感器展示了宽线性范围（10 ng mL至50 fg mL），检测限极低（5 fg mL）。这种CKMB免疫传感器还对人血清中常见的干扰物表现出显著的选择性、重现性、稳定性和抗干扰能力。此外，该免疫传感器在用于临床诊断的实际血清样本检测方面具有巨大潜力。
