Center for the Study of Venoms and Venomous Animals (CEVAP), São Paulo State University (UNESP-Univ Estadual Paulista), Botucatu, São Paulo, Brazil.
Immunology Center, Adolfo Lutz Institute, São Paulo, São Paulo, Brazil.
Front Immunol. 2024 Jan 23;14:1303353. doi: 10.3389/fimmu.2023.1303353. eCollection 2023.
Since their discovery in the 1990s, heavy chain antibodies have garnered significant interest in the scientific community. These antibodies, found in camelids such as llamas and alpacas, exhibit distinct characteristics from conventional antibodies due to the absence of a light chain in their structure. Furthermore, they possess a single antigen-binding domain known as VHH or Nanobody (Nb). With a small size of approximately 15 kDa, these Nbs demonstrate improved characteristics compared to conventional antibodies, including greater physicochemical stability and enhanced biodistribution, enabling them to bind inaccessible epitopes more effectively. As a result, Nbs have found numerous applications in various medical and veterinary fields, particularly in diagnostics and therapeutics. Advances in biotechnology have made the production of recombinant antibodies feasible and compatible with large-scale manufacturing. Through the construction of immune phage libraries that display VHHs and subsequent selection through biopanning, it has become possible to isolate specific Nbs targeting pharmaceutical targets of interest, such as viruses. This review describes the processes involved in nanobody production, from hyperimmunization to purification, with the aim of their application in the pharmaceutical industry.
自 20 世纪 90 年代发现以来,重链抗体在科学界引起了广泛关注。这些抗体存在于骆驼科动物(如羊驼和骆马)中,由于其结构中缺少轻链,因此具有与传统抗体不同的特性。此外,它们具有单一的抗原结合结构域,称为 VHH 或纳米抗体(Nb)。这些 Nb 的分子量约为 15 kDa,与传统抗体相比具有更好的特性,包括更高的物理化学稳定性和增强的生物分布,使其能够更有效地结合难以接近的表位。因此,Nb 在医学和兽医领域的各种应用中得到了广泛应用,特别是在诊断和治疗方面。生物技术的进步使得重组抗体的生产变得可行,并与大规模制造兼容。通过构建展示 VHH 的免疫噬菌体文库,并通过生物淘选进行选择,可以分离出针对感兴趣的药物靶点(如病毒)的特异性 Nb。本综述描述了从免疫接种到纯化的纳米抗体生产过程,旨在将其应用于制药行业。