Fan Xiying, Gong Yibo, Liu Wenshuai, Lam Kong Peng, Xu Shengli, Xue Song, Nian Rui
CAS Key Laboratory of Biobased Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, No. 189, Songling Road, Qingdao, 266101, China.
Shandong Energy Institute, No. 189, Songling Road, Qingdao, 266101, China.
Chemistry. 2025 Aug 13;31(45):e04431. doi: 10.1002/chem.202404431. Epub 2025 Jul 24.
Natural toxins in food pose significant health risks, such as cancer, deformities, etc., demanding efficient and accurate surveillance techniques. Immunoassays become reliable alternatives to traditional chromatographic technologies for rapid, routine, and large-scale detection of natural toxins, but their performance depends highly on antibodies used, which are limited by stability issues and productions challenge. Nanobodies (Nbs) overcome these limitations, enhancing immunoassay sensitivity, specificity, and practicality for low-abundance toxin detection due to their good penetration/binding ability, inherent stability, and scalable production. This review offers a comprehensive, bottom-to-top overview of emerging Nb-based immunoassays, from foundational Nbs to advanced applications in natural toxin detection. The bioactivity of Nbs was summarized, focusing on their structural and functional characteristics critical for advancing immunoassays. Subsequently, the advanced methods for their screening, expression, and purification were outlined. Based on the natural toxins they target, both conventional and innovative Nb-based immunoassays are reviewed, in which corresponding Nbs modification strategies are highlighted for improving the affinity and sensitivity. The current challenges, potential strategies, and future opportunities for developing and applying Nb-based immunoassays are also discussed in natural toxin detection. This review, bridging fundamental Nb advances and immunoassay innovation, offers valuable perspectives to guide future development of Nb-based detection platforms.
食品中的天然毒素会带来重大健康风险，如癌症、畸形等，这就需要高效且准确的监测技术。免疫分析成为传统色谱技术的可靠替代方法，可用于快速、常规且大规模地检测天然毒素，但其性能很大程度上取决于所使用的抗体，而抗体受稳定性问题和生产挑战的限制。纳米抗体（Nbs）克服了这些局限性，由于其良好的穿透/结合能力、固有稳定性和可扩展生产性，增强了免疫分析对低丰度毒素检测的灵敏度、特异性和实用性。本综述全面且自下而上地概述了新兴的基于纳米抗体的免疫分析，从基础的纳米抗体到在天然毒素检测中的先进应用。总结了纳米抗体的生物活性，重点关注其对推进免疫分析至关重要的结构和功能特征。随后，概述了其筛选、表达和纯化的先进方法。基于它们所靶向的天然毒素，对传统和创新的基于纳米抗体的免疫分析进行了综述，其中突出了相应的纳米抗体修饰策略以提高亲和力和灵敏度。在天然毒素检测中还讨论了开发和应用基于纳米抗体的免疫分析的当前挑战、潜在策略和未来机遇。本综述将纳米抗体的基础进展与免疫分析创新联系起来，为指导基于纳米抗体的检测平台的未来发展提供了有价值的观点。
