Department of Biochemistry, Faculty of Science, Ege University, Bornova, Izmir 35100, Turkey.
Department of Chemistry, Faculty of Science, Ege University, Bornova, Izmir 35100, Turkey.
Biosensors (Basel). 2023 Jun 23;13(7):673. doi: 10.3390/bios13070673.
Diagnostic techniques based on biomolecules have application potential that can be realized in many fields, such as disease diagnosis, bioprocess imaging, food/beverage industries, and environmental pollutant imaging. Successful surface immobilization of biomolecules is critical to increasing the stabilization, sensitivity, and selectivity of biomolecules used in bioassay systems. Nanofibers are good candidates for the immobilization of biomolecules owing to many advantages such as morphology and pore size. In this study, montmorillonite (MMT) clay is modified with poly(amidoamine) (PAMAM) generation 3 (PAMAM) and added to polystyrene (PS) solutions, following which PS/MMT-PAMAM nanofibers are obtained using the electrospinning method. The nanofibers are obtained by testing PS% (wt%) and MMT-PAMAM% (wt%) ratios and characterized with scanning electron microscopy. Antiserum amyloid A antibody (Anti-SAA) is then conjugated to the nanofibers on the electrode surface via covalent bonds using a zero-length cross linker. Finally, the obtained selective surface is used for electrochemical determination of serum amyloid A (SAA) levels. The linear range of PS/MMT-PAMAM/Anti-SAA is between 1 and 200 ng/mL SAA, and the detection limit is 0.57 ng/mL SAA. The applicability of PS/MMT-PAMAM/Anti-SAA is investigated by taking measurements in synthetic saliva and serum both containing SAA.
基于生物分子的诊断技术具有很大的应用潜力,可以在许多领域得到应用,如疾病诊断、生物过程成像、食品/饮料工业和环境污染物成像。成功地将生物分子固定在表面上对于提高生物分析系统中生物分子的稳定性、灵敏度和选择性至关重要。由于形态和孔径等诸多优点,纳米纤维是固定生物分子的良好候选材料。在这项研究中,蒙脱土(MMT)粘土用聚(酰胺-胺)(PAMAM)第 3 代(PAMAM)进行修饰,并添加到聚苯乙烯(PS)溶液中,然后通过静电纺丝法获得 PS/MMT-PAMAM 纳米纤维。通过测试 PS%(wt%)和 MMT-PAMAM%(wt%)的比例获得纳米纤维,并通过扫描电子显微镜进行表征。然后通过使用零长度交联剂通过共价键将抗血清淀粉样蛋白 A 抗体(Anti-SAA)偶联到电极表面上的纳米纤维上。最后,获得的选择性表面用于电化学测定血清淀粉样蛋白 A(SAA)水平。PS/MMT-PAMAM/Anti-SAA 的线性范围在 1 到 200ng/mL SAA 之间,检测限为 0.57ng/mL SAA。通过在含有 SAA 的合成唾液和血清中进行测量,研究了 PS/MMT-PAMAM/Anti-SAA 的适用性。