Programa de Pós-Graduação em Microbiologia, Parasitologia e Patologia - Departamento de Patologia Básica, Universidade Federal do Paraná, CEP 81531-980 Curitiba, PR, Brazil; ISP UMR 1282, INRA, Université de Tours, Team BioMAP, 31 avenue Monge, 37200 Tours, France.
Programa de Pós-Graduação em Microbiologia, Parasitologia e Patologia - Departamento de Patologia Básica, Universidade Federal do Paraná, CEP 81531-980 Curitiba, PR, Brazil.
Int J Biol Macromol. 2022 Sep 1;216:465-474. doi: 10.1016/j.ijbiomac.2022.06.195. Epub 2022 Jul 6.
The use of monoclonal antibodies (mAbs) in therapy is gradually advancing and discussions entail its safety, rentability and effectiveness. To this date, around a hundred mAbs have been approved by the FDA for the treatment of various diseases. Aiming for their large-scale production, recombinant DNA technology is mainly employed, and antibodies can be expressed in various eukaryotic and prokaryotic systems. Moreover, considering their heterologous origin and potential immunogenicity, various strategies have been developed for mAb humanization, considering that around 50 % of commercial mAbs are humanized. Hence, we introduce LimAb7, a mouse mAb capable of binding and neutralizing brown spider's Loxosceles intermedia dermonecrotic toxins in vivo/in vitro. This antibody has been produced in mouse and humanized scFv and diabody formats, however results indicated losses in antigen-binding affinity, stability, and neutralizing ability. Intending to develop evolved, stable, and neutralizing antibody fragments, we report for the first time the design of humanized antibody V-domains produced as Fab fragments, against spider venom toxins. Improvements in constructs were observed regarding their physicochemical stability, target binding and binding pattern maintenance. As their neutralizing features remain to be characterized, we believe this data sheds new light on antibody humanization by producing a parental molecule in different recombinant formats.
单克隆抗体（mAbs）在治疗中的应用正在逐步推进，人们对其安全性、盈利性和有效性进行了讨论。迄今为止，FDA 已经批准了大约 100 种 mAbs 用于治疗各种疾病。为了实现大规模生产，主要采用重组 DNA 技术，可以在各种真核和原核系统中表达抗体。此外，考虑到它们的异源性和潜在免疫原性，已经开发了各种 mAb 人源化策略，因为大约 50%的商业 mAbs 已经人源化。因此，我们介绍了 LimAb7，这是一种能够在体内/体外结合和中和棕色蜘蛛 Loxosceles intermedia 皮肤坏死毒素的鼠源 mAb。该抗体已在小鼠和人源化 scFv 和二价抗体形式中生产，但结果表明抗原结合亲和力、稳定性和中和能力都有所下降。为了开发进化稳定且具有中和能力的抗体片段，我们首次报告了针对蜘蛛毒液毒素的 Fab 片段形式产生的人源化抗体 V 结构域的设计。在构建体方面观察到了一些改进，包括其物理化学稳定性、靶标结合和结合模式保持。由于其中和特性仍有待表征，我们认为通过以不同重组形式产生亲本分子，为抗体人源化提供了新的思路。
