BioMark/CINTESIS@ISEP, Instituto Superior de Engenharia do Instituto Politécnico do Porto, 4249-015 Porto, Portugal.
Departamento de Ciências Médicas, Universidade de Aveiro, 3810-193 Aveiro, Portugal.
Biosensors (Basel). 2018 Jan 19;8(1):8. doi: 10.3390/bios8010008.
This work reports a very simple approach for creating a synthetic antibody against any protein of interest and its application in potentiometric transduction. The selected protein was Breast Cancer Antigen (CA 15-3), which is implicated in breast cancer disease and used to follow-up breast cancer patients during treatment. The new material with antibody-like properties was obtained by molecular-imprinting technology, prepared by electropolymerizing pyrrol (Py, 5.0 × 10 mol/L) around Breast Cancer Antigen (CA 15-3) (100 U/mL) on a fluorine doped tin oxide (FTO) conductive glass support. Cyclic voltammetry was employed for this purpose. All solutions were prepared in 4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer, of pH 6.5. The biomarker was removed from the imprinted sites by chemical action of ethanol. The biomimetic material was then included in poly vinyl chloride (PVC) plasticized membranes to act as potentiometric ionophore, having or not a lipophilic ionic additive added. The corresponding selective electrodes were evaluated by calibration curves (in buffer and in synthetic serum) and by selectivity testing. The best analytical performance was obtained by selective electrodes including the plastic antibody and no lipophilic additive. The average limits of detection were 1.07 U/mL of CA 15-3, with a linear response from 1.44 to 13.2 U/mL and a cationic slope of 44.5 mV/decade. Overall, the lipophilic additives yielded no advantage to the overall potentiometric performance. The application of the MIP-based electrodes to the analysis of spiked synthetic serum showed precise and accurate results.
这项工作报道了一种非常简单的方法,用于创建针对任何感兴趣的蛋白质的合成抗体,并将其应用于电位转换中。选择的蛋白质是乳腺癌抗原(CA 15-3),它与乳腺癌疾病有关,用于在治疗期间随访乳腺癌患者。具有类似抗体特性的新材料是通过分子印迹技术获得的,该技术通过在氟掺杂氧化锡(FTO)导电玻璃衬底上围绕乳腺癌抗原(CA 15-3)(100 U/mL)电聚合吡咯(Py,5.0×10 mol/L)来制备。为此目的使用了循环伏安法。所有溶液均在 4-(2-羟乙基)-1-哌嗪乙磺酸(HEPES)缓冲液中制备,pH 值为 6.5。通过乙醇的化学作用从印迹部位去除生物标志物。然后将仿生材料包含在聚氯乙烯(PVC)增塑膜中作为电位离子载体,具有或不具有添加的亲脂性离子添加剂。通过校准曲线(在缓冲液和合成血清中)和选择性测试评估相应的选择性电极。通过包括塑料抗体和没有亲脂性添加剂的选择性电极获得最佳的分析性能。平均检出限为 CA 15-3 的 1.07 U/mL,线性响应范围为 1.44 至 13.2 U/mL,阳离子斜率为 44.5 mV/decade。总体而言,亲脂性添加剂对整体电位性能没有优势。基于 MIP 的电极在分析加标合成血清中的应用显示出精确和准确的结果。