Erkal-Aytemur Aslı, Mülazımoğlu İbrahim Ender, Üstündağ Zafer, Caglayan Mustafa Oguzhan
Alanya Alaaddin Keykubat University, R.K. Faculty of Engineering, Fundamental Science, Antalya, Turkey.
Necmettin Erbakan University, A.K. Education Faculty, Chemistry Department, Konya, Turkey.
Talanta. 2024 Sep 1;277:126376. doi: 10.1016/j.talanta.2024.126376. Epub 2024 Jun 9.
In this study, a quartz crystal microbalance (QCM) aptasensor for carcinoembryonic antigen (CEA), a well-known biomarker for various cancer types, was reported, utilizing two different aptamers. To achieve this, a nanofilm of 4-mercaptophenyl was electrochemically attached to gold-coated QCM crystal surfaces via the reduction of 4-mercaptobenzenediazonium salt (4 MB-DAT) using cyclic voltammetry. Subsequently, gold nanoparticles (AuNP) were affixed to this structure, and then aptamers (antiCEA1 and antiCEA2) modified with SH-functional ends bound to AuNPs completed the modification. The analytical performance of the CEA sensor was evaluated through simultaneous QCM measurements employing CEA solutions ranging from 0.1 ng/mL to 25 ng/mL. The detection limit (LOD) for CEA was determined to be 102 pg/mL for antiCEA1 and 108 pg/mL for antiCEA2 aptamers. Interday and intraday precision and accuracy tests yielded maximum results of 4.3 and + 3.8, respectively, for both aptasensors, as measured by relative standard deviation (RSD%) and relative error (RE%). The kinetic data of the aptasensors resulted in affinity values (K) of 0.43 ± 0.14 nM for antiCEA1 and 0.75 ± 0.42 nM for antiCEA2. These values were lower than the reported values of 3.9 nM and 37.8 nM for both aptamers, respectively. The selectivity of the aptasensor was evaluated by measuring the signal changes caused by alpha-fetoprotein (AFP), cancer antigen (CA-125), and vascular endothelial growth factor (VEGF-165) individually and together at a concentration of 500 ng/mL, resulting in a maximum 4.1 % change, which was comparable to precision and accuracy values reported in the literature. After confirming the selectivity of the aptamers, recovery experiments were conducted using spiked commercial serum samples to simulate real samples, and the lowest recovery value obtained was 95.4 %. It was determined that two different aptasensors could be successfully used for the QCM-based detection of CEA in this study.
在本研究中，报道了一种用于癌胚抗原（CEA）的石英晶体微天平（QCM）适配体传感器，CEA是一种用于多种癌症类型的著名生物标志物，该传感器利用了两种不同的适配体。为此，通过循环伏安法还原4-巯基苯重氮盐（4-MB-DAT），将4-巯基苯纳米膜电化学附着在涂金的QCM晶体表面。随后，将金纳米颗粒（AuNP）固定在该结构上，然后用与AuNP结合的SH功能端修饰的适配体（抗CEA1和抗CEA2）完成修饰。通过使用浓度范围为0.1 ng/mL至25 ng/mL的CEA溶液进行同步QCM测量，评估了CEA传感器的分析性能。抗CEA1适配体对CEA的检测限（LOD）测定为102 pg/mL，抗CEA2适配体的检测限为108 pg/mL。日间和日内精密度和准确度测试结果表明，两种适配体传感器通过相对标准偏差（RSD%）和相对误差（RE%）测量，最大值分别为4.3和+3.8。适配体传感器的动力学数据得出抗CEA1的亲和值（K）为0.43±0.14 nM，抗CEA2的亲和值为0.75±0.42 nM。这些值分别低于两种适配体报道的3.9 nM和37.8 nM的值。通过分别测量以及一起测量浓度为500 ng/mL的甲胎蛋白（AFP）、癌抗原（CA-125）和血管内皮生长因子（VEGF-165）引起的信号变化，评估了适配体传感器的选择性，信号变化最大值为4.1%，这与文献报道的精密度和准确度值相当。在确认适配体的选择性后，使用加标的商业血清样本进行回收实验以模拟实际样本，获得的最低回收率为95.4%。本研究确定两种不同的适配体传感器可成功用于基于QCM的CEA检测。
