Human Monoclonal Antibodies that Bind Botulinum Toxins

描述（由申请人提供）：肉毒杆菌神经毒素（BoNT）是已知的最有效的生物毒素，是麻痹性疾病、肉毒杆菌中毒的原因，也是潜在的生物恐怖武器。用抗体治疗剂进行被动免疫被认为是BoNT暴露的理想对策，其中最安全的是由人单克隆抗体组成。产生BoNT的有效抗体治疗策略的一个重要障碍是BoNT存在于至少7种血清型中，并且迄今为止克隆的大多数抗体不能结合一种以上的血清型。此外，有效的中和需要使用抗体的三联体组合，使得3种毒素血清型的中和将需要至少9种不同的抗体。该项目通过探索一个平台来解决这个问题，该平台用于创建能够结合和中和BoNT、A、B和E三种不同血清型的人单克隆抗体治疗剂。该计划的基础是一种克隆人抗体的杂交瘤方法，该方法已经产生了许多交叉反应抗体。这与使用融合肽增加BoNT抗体的体内效力的新方法相结合，所述融合肽可以增加BoNT从血流中的清除速率。该研究的一个重要方面是使用人单克隆抗体开发的超高灵敏度ELISA，其将允许在真实世界的体内暴露场景中对结合抗体的低剂量BoNT的药代动力学研究：融合肽复合物。 公共卫生相关性：肉毒杆菌神经毒素（BoNT）已被疾病控制中心列为A类生物恐怖剂。多血清型特异性人类抗体的产生和研究，结合增加血液中BoNT清除的新技术，将有助于为肉毒杆菌中毒或BoNT生物恐怖袭击创造综合治疗策略。它还将提供一个广泛的理解，如何类似的方法可以用来治疗各种传染病和毒素暴露。

DESCRIPTION (provided by applicant): Botulinum neurotoxin (BoNT) is the most potent biological toxin known, the cause of the paralytic disease, botulism, and a potential bioterrorist weapon. Passive immunization with antibody therapeutics is considered to be the ideal countermeasure for a BoNT exposure, and the safest of these would be composed of human monoclonal antibodies. An important obstacle to the production of an effective antibody therapeutic strategy for BoNT is that BoNT exists in at least 7 serotypes, and most of the antibodies cloned thus far are not able to bind more than one serotype. In addition, effective neutralization requires that triplet combinations of antibodies be employed, so that neutralization of 3 toxin serotypes would require at least 9 distinct antibodies. This project addresses this problem by exploring a platform for the creation of human monoclonal antibody therapeutics capable of binding and neutralizing three distinct serotypes of BoNT, A, B, and E. The foundation of the plan is a hybridoma method of human antibody cloning that has generated a number of cross-reactive antibodies. This is combined with a novel means of increasing the in vivo potency of BoNT antibodies using a fusion peptide that may increase the rate of clearance of BoNT from the bloodstream. An important aspect of the study is an ultra-high sensitivity ELISA, developed using human monoclonal antibodies, which will allow pharmacokinetic study of low doses of BoNT bound to antibody:fusion peptide complexes in real-world in vivo exposure scenarios. PUBLIC HEALTH RELEVANCE: Botulinum neurotoxin (BoNT) has been listed as a Category A Select Bioterror agent by the Centers for Disease Control. Generation and study of multi-serotype-specific human antibodies, in combination with a novel technology for increasing BoNT clearance from the bloodstream, will help in the creation of a comprehensive therapeutic strategy for botulism or a BoNT bioterror attack. It will also provide a broad understanding of how similar approaches could be used to treat a variety of infectious diseases and toxin exposures.