An Electrochemical Immuno-cytosensor Modified with Nanofibers for the Determination of a Carcinoembryonic Antigen

IF 5.5 3区 工程技术 Q1 BIOCHEMICAL RESEARCH METHODS
Zehra Yildizbakan, Derya Bal Altuntaş, Hatice Sevim Nalkiran, Sema Aslan, Atilla Eren Mamuk, Çağdaş Koçak, Şeymanur Kurt, İhsan Nalkiran, Çiğdem Yener, C. Gökhan Ünlü
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引用次数: 0

Abstract

In this study, La0.25Fe0.75FeO3 (PNp)perovskite nanoparticle was synthesized using the sol–gel method. PNp-coated polyacrylonitrile (PAN) nanofibers were prepared by electrospinning on the pencil graphite electrode (PGE) surface. In another step, carcinoembryonic antigen (CEA) was loaded with CEA antibodies (Anti-CEA) as a biomarker receptor. Finally, PGE/PAN@PNp/Anti-CEA was used for CEA detection. Optimization steps and cell culture steps were performed using differential pulse voltammetry (DPV). The use of this composite system is a novel immunosensor development approach for label-free detection of CEA. Under optimum conditions, detection limit (LOD) of PGE/PAN@PNp/Anti-CEA immunosensor LOD 1.48 ng/mL, limit of quantification (LOQ) = 4.94 ng/mL, reproducibility 1.46% (n = 5) and R2 = 0.9984 for antigen concentration within a linear working range of 0.1–10 ng/mL. Also, immunosensor recovery in real serum samples containing dopamine and ascorbic acid was found as 98.94 ± 7.43. It has great potential in clinical screening of different cancer biomarkers. The number of cells attached to the PGE/PAN@PNp/Anti-CEA/BSA(bovine serum)/CEA surface decreased in RT-4(bladder cancer), MDA-MB-231 (triple-negative breast adenocarcinoma cell line), and T98G cells (glioblastoma multiforme cell line), which are known as CEA-negative cell lines, whereas the number of MCF-7 cells (estrogen-sensitive human breast cancer cell line, known to be CEA positive) attached to the PGE/PAN@PNp/Anti-CEA/BSA/CEA surface increased, indicating higher affinity to the immunosensor surface. As a result, while MCF-7, which is CEA positive, can be determined best when using an immune-cytosensor, the cell that can be best determined with cytosensors was found to be RT-4.

Abstract Image

用纳米纤维修饰的电化学免疫细胞传感器用于检测癌胚抗原
本研究采用溶胶-凝胶法合成了 La0.25Fe0.75FeO3 (PNp) 包晶纳米粒子。在铅笔石墨电极(PGE)表面通过电纺丝制备了 PNp 涂层聚丙烯腈(PAN)纳米纤维。在另一个步骤中,将癌胚抗原(CEA)与 CEA 抗体(Anti-CEA)负载在一起,作为生物标记受体。最后,PGE/PAN@PNp/Anti-CEA 被用于检测 CEA。使用差分脉冲伏安法(DPV)进行了优化步骤和细胞培养步骤。使用这种复合系统是一种新型免疫传感器开发方法,可用于无标记检测 CEA。在最佳条件下,PGE/PAN@PNp/抗 CEA 免疫传感器的检测限(LOD)为 1.48 纳克/毫升,定量限(LOQ)为 4.94 纳克/毫升,重现性为 1.46%(n = 5),抗原浓度在 0.1-10 纳克/毫升线性工作范围内的 R2 = 0.9984。此外,在含有多巴胺和抗坏血酸的真实血清样本中,免疫传感器的回收率为 98.94 ± 7.43。它在不同癌症生物标记物的临床筛选方面具有巨大潜力。在 RT-4(膀胱癌)、MDA-MB-231(三阴性乳腺癌细胞系)和 T98G 细胞(多形性胶质母细胞瘤细胞系)中,附着在 PGE/PAN@PNp/Anti-CEA/BSA (牛血清)/CEA 表面的细胞数量减少、而附着在 PGE/PAN@PNp/Anti-CEA/BSA/CEA 表面的 MCF-7 细胞(对雌激素敏感的人类乳腺癌细胞系,已知为 CEA 阳性)的数量却增加了,这表明它们与免疫传感器表面的亲和力提高了。因此,虽然使用免疫细胞传感器能最好地测定 CEA 阳性的 MCF-7,但发现使用细胞传感器能最好地测定的细胞是 RT-4。
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来源期刊
BioChip Journal
BioChip Journal 生物-生化研究方法
CiteScore
7.70
自引率
16.30%
发文量
47
审稿时长
6-12 weeks
期刊介绍: BioChip Journal publishes original research and reviews in all areas of the biochip technology in the following disciplines, including protein chip, DNA chip, cell chip, lab-on-a-chip, bio-MEMS, biosensor, micro/nano mechanics, microfluidics, high-throughput screening technology, medical science, genomics, proteomics, bioinformatics, medical diagnostics, environmental monitoring and micro/nanotechnology. The Journal is committed to rapid peer review to ensure the publication of highest quality original research and timely news and review articles.
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