Tang Yujiao, Zhao Xiaomin, Nie Zhenlin, He Bangshun, Deng Dawei
Department of Biomedical Engineering, School of Engineering, China Pharmaceutical University, Nanjing 211198, China.
Department of Pharmaceutical Engineering, School of Engineering, China Pharmaceutical University, Nanjing 211198, China.
ACS Appl Bio Mater. 2025 Jul 21;8(7):6252-6260. doi: 10.1021/acsabm.5c00745. Epub 2025 Jun 28.
The monoclonal antibody (mAb) used in a traditional Lateral Flow Immunoassay (LFIA) has many limitations, particularly related to its poor affinity, specificity, and stability. In this study, a nanobody (VHH) was developed with improved tolerability and higher specificity for the target. The strong binding affinity between VHH and survivin was emphasized by simulated docking, demonstrating the feasibility and effectiveness of replacing mAb in the detection of urinary survivin based on LFIA. In addition, the superior performance of VHH compared with mAb under extreme conditions was highlighted. To further enhance the detection performance, VHH was conjugated with an Au nanoparticle cluster (AuNP cluster). The pore size structure of the AuNP cluster led to an increased abundance of AuNPs, which amplified the signal while providing adequate adsorption sites for VHH coupling, thereby achieving a synergistic enhancement in the sensitivity and specificity of the AuNP cluster-VHH probe. Consequently, the detection limit for survivin was established at 3.90 ng/mL, facilitating noninvasive detection and monitoring in early-stage bladder cancer and during post-treatment surveillance. This insight underscores the key role of VHH in enhancing the performance of LFIA and the potential of developing a promising detection tool based on urinary biomarkers.
传统侧向流动免疫分析（LFIA）中使用的单克隆抗体（mAb）有许多局限性，特别是其亲和力、特异性和稳定性较差。在本研究中，开发了一种对目标具有更高耐受性和更高特异性的纳米抗体（VHH）。通过模拟对接强调了VHH与生存素之间的强结合亲和力，证明了在基于LFIA的尿液生存素检测中替代mAb的可行性和有效性。此外，还突出了VHH在极端条件下与mAb相比的优越性能。为了进一步提高检测性能，将VHH与金纳米颗粒簇（AuNP簇）偶联。AuNP簇的孔径结构导致AuNP丰度增加，在放大信号的同时为VHH偶联提供了足够的吸附位点，从而实现了AuNP簇-VHH探针灵敏度和特异性的协同增强。因此，生存素的检测限确定为3.90 ng/mL，便于在早期膀胱癌及治疗后监测中进行无创检测和监测。这一见解强调了VHH在提高LFIA性能方面的关键作用以及基于尿液生物标志物开发有前景的检测工具的潜力。
