Al-Seragi Mohammed, Chen Yilun, Duong van Hoa Franck
Department of Biochemistry and Molecular Biology, Life Sciences Institute, University of British Columbia, Vancouver, British Columbia, Canada.
Biochem Soc Trans. 2025 Feb 7;53(1):BST20241419. doi: 10.1042/BST20241419.
NANOBODIES® (Nbs) have emerged as valuable tools across therapeutic, diagnostic, and industrial applications owing to their small size and consequent ability to bind unique epitopes inaccessible to conventional antibodies. While Nbs retrieved from immune libraries normally possess sufficient affinity and specificity for their cognate antigens in the practical use case, their multimerization will often increase functional affinity via avidity effects. Therefore, to rescue binding affinity and broaden targeting specificities, recent efforts have focused on conjugating multiple Nb clones - of identical or unique antigen cognates - together. In vivo and in vitro approaches, including flexible linkers, antibody domains, self-assembling coiled coils, chemical conjugation, and self-clustering hydrophobic sequences, have been employed to produce multivalent and multispecific Nb constructs. Examples of successful Nb multimerization are diverse, ranging from immunoassaying reagents to virus-neutralizing moieties. This review aims to recapitulate the in vivo and in vitro modalities to produce multivalent and multispecific Nbs while highlighting the applications, advantages, and drawbacks tied to each method.
纳米抗体（Nbs）因其体积小，因而能够结合传统抗体无法触及的独特表位，已成为治疗、诊断和工业应用中的重要工具。虽然从免疫文库中获得的纳米抗体在实际应用中通常对其同源抗原具有足够的亲和力和特异性，但它们的多聚化往往会通过亲合力效应增加功能亲和力。因此，为了挽救结合亲和力并拓宽靶向特异性，最近的研究重点是将多个相同或独特抗原同源的纳米抗体克隆连接在一起。体内和体外方法，包括柔性接头、抗体结构域、自组装卷曲螺旋、化学偶联和自聚集疏水序列，已被用于生产多价和多特异性纳米抗体构建体。成功的纳米抗体多聚化实例多种多样，从免疫分析试剂到病毒中和部分。本综述旨在概括体内和体外产生多价和多特异性纳米抗体的方法，同时强调每种方法的应用、优点和缺点。
