Institut Pasteur, Unité des Bactéries Anaérobies et Toxines, 25 Avenue du Docteur Roux, 75015 Paris, France.
Institut de Recherche Biomédicale des Armées (IRBA-CRSSA), Département de Microbiologie, Unité de Biotechnologie des Anticorps et Des Toxins, Cedex 38702 La Tronche, France.
Toxins (Basel). 2017 Oct 2;9(10):309. doi: 10.3390/toxins9100309.
The goal of the AntiBotABE Program was the development of recombinant antibodies that neutralize botulinum neurotoxins (BoNT) A, B and E. These serotypes are lethal and responsible for most human botulinum cases. To improve therapeutic efficacy, the heavy and light chains (HC and LC) of the three BoNT serotypes were targeted to achieve a synergistic effect (oligoclonal antibodies). For antibody isolation, macaques were immunized with the recombinant and non-toxic BoNT/A, B or E, HC or LC, followed by the generation of immune phage-display libraries. Antibodies were selected from these libraries against the holotoxin and further analyzed in in vitro and ex vivo assays. For each library, the best ex vivo neutralizing antibody fragments were germline-humanized and expressed as immunoglobulin G (IgGs). The IgGs were tested in vivo, in a standardized model of protection, and challenged with toxins obtained from collections of strains. Protective antibody combinations against BoNT/A and BoNT/B were evidenced and for BoNT/E, the anti-LC antibody alone was found highly protective. The combination of these five antibodies as an oligoclonal antibody cocktail can be clinically and regulatorily developed while their high "humanness" predicts a high tolerance in humans.
抗 Botulinum 抗体计划的目标是开发能够中和肉毒梭菌神经毒素（BoNT）A、B 和 E 的重组抗体。这些血清型是致命的，也是大多数人类肉毒中毒病例的罪魁祸首。为了提高治疗效果，针对三种 BoNT 血清型的重链（HC）和轻链（LC）进行了靶向，以实现协同效应（多克隆抗体）。为了进行抗体分离，猕猴用重组和无毒的 BoNT/A、B 或 E、HC 或 LC 进行免疫，随后生成免疫噬菌体展示文库。从这些文库中选择针对全毒素的抗体，并在体外和体内进行进一步分析。对于每个文库，最佳的体外中和抗体片段进行种系人源化，并表达为免疫球蛋白 G（IgG）。在标准化的保护模型中对 IgG 进行体内测试，并使用从菌株收集物中获得的毒素进行挑战。针对 BoNT/A 和 BoNT/B 证明了保护性抗体组合的存在，对于 BoNT/E，单独的抗 LC 抗体被发现具有高度的保护作用。将这五种抗体作为多克隆抗体鸡尾酒进行临床和监管开发，同时它们的高“人源化”可预测在人类中具有较高的耐受性。
