Nanobiotechnology Research Group, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia; Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
Nanobiotechnology Research Group, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia; Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
Bioelectrochemistry. 2022 Feb;143:107952. doi: 10.1016/j.bioelechem.2021.107952. Epub 2021 Sep 20.
The diagnosis of hepatitis B virus (HBV) and monitoring of the vaccination efficiency against HBV require real-time analysis. The presence of antibody against hepatitis B virus surface antigen (anti-HBsAg) as a result of HBV infection and/or immunization may indicate individual immune status towards HBV. This study investigated the ability of a bio-nanogate-based displacement immunosensing strategy in detecting anti-HBsAg antibody, via nonspecific-binding between polyamidoamine dendrimers encapsulated gold nanoparticles (PAMAM-Au) and the 'antigenic determinant' region (aD) of HBsAg. For this purpose, maltose binding protein harbouring the aD region (MBP-aD) was synthesized as a bioreceptor and immobilized on the screen-printed carbon electrode (SPCE). Following that, PAMAM-Au was deposited on MBP-aD, forming the 'gate' and was used as a monitoring agent. Under optimal conditions, the high specificity of anti-HBsAg antibody towards MBP-aD displaced PAMAM-Au causing the decrement of anodic peak in differential pulse voltammetry (DPV) analysis. The signal changes were proportionally related to the concentration of anti-HBsAg antibody, in a range of 1 - 1000 mIU/mL with a limit of detection (LOD) of 2.5 mIU/mL. The results also showed high specificity and selectivity of the immunosensor platform in detecting anti-HBsAg antibody both in spiked buffer and human serum samples.
乙型肝炎病毒 (HBV) 的诊断和 HBV 疫苗接种效率的监测需要实时分析。由于 HBV 感染和/或免疫而产生的乙型肝炎病毒表面抗原抗体 (抗-HBsAg) 的存在可能表明个体对 HBV 的免疫状态。本研究通过聚酰胺胺树枝状大分子包裹的金纳米粒子 (PAMAM-Au) 与 HBsAg 的“抗原决定簇”(aD) 之间的非特异性结合，研究了基于生物纳米门控的置换免疫传感策略检测抗-HBsAg 抗体的能力。为此，合成了含有 aD 区的麦芽糖结合蛋白 (MBP-aD) 作为生物受体并固定在丝网印刷碳电极 (SPCE) 上。随后，将 PAMAM-Au 沉积在 MBP-aD 上，形成“门”并用作监测剂。在最佳条件下，抗-HBsAg 抗体对 MBP-aD 的高特异性会置换 PAMAM-Au，导致差分脉冲伏安法 (DPV) 分析中阳极峰的减小。信号变化与抗-HBsAg 抗体的浓度呈比例关系，在 1 - 1000 mIU/mL 的范围内，检测限 (LOD) 为 2.5 mIU/mL。结果还表明，该免疫传感器平台在检测缓冲液和人血清样本中的抗-HBsAg 抗体时具有高特异性和选择性。
