Human Monoclonal Antibodies that Bind Botulinum Toxins

说明(申请人提供)：肉毒杆菌神经毒素(BONT)是一种非常有效的神经肌肉阻滞剂，由于其极端的杀伤力和环境来源的可获得性，已被美国政府指定为A类精选生物恐怖制剂。目前针对BONT暴露的对策是不够的，迫切需要能够抵消BONT暴露影响的治疗性抗体。我们的工作假设是，结合BONT的人类抗体组合将有效地保护个人免受BONT暴露的并发症的影响。我们最近开发了一种新的杂交瘤方案来克隆人抗体，并用它来克隆结合A型和B型BoNT的抗体。其中一种A型抗体在体内是中和的。我们计划使用我们的方法来克隆结合和中和A型和B型肉毒毒素的天然人类抗体。为此，我们将从接种肉毒毒素疫苗的人身上获得外周血淋巴细胞。我们将单独和联合评估克隆抗体的体外和体内中和活性。我们将确定抗体的结合和初级序列的特征，以确定与中和毒素的能力相关的特征，并在中和毒素方面协同合作。这些实验的主要结果将是创造出克隆的人类抗体组合，可以有效地抵消BONT/A和BONT/B的暴露。

DESCRIPTION (provided by applicant): Botulinum neurotoxin (BoNT) is an extremely potent neuromuscular blocker that has been designated a category A select bioterror agent by the United States Government because of its extreme lethality and its availability from environmental sources. Current countermeasures for BoNT exposure are insufficient, and there is an urgent need for therapeutic antibodies able to counteract the effects of BoNT exposure. Our working hypothesis is that a combination of human antibodies that bind BoNT will potently protect individuals from the complications of BoNT exposure. We have recently developed a novel hybridoma protocol for the cloning of human antibodies and have used it to clone antibodies that bind BoNT serotypes A and B. One of the serotype A antibodies is neutralizing in vivo. We plan to use our method to clone native human antibodies that bind and neutralize botulinum toxin serotypes A and B. For this purpose we will obtain peripheral blood lymphocytes from individuals vaccinated with the botulinum toxoid vaccine. We will assess in vitro and in vivo neutralizing activities of the cloned antibodies, singly and in combination. We will characterize the binding and primary sequences of the antibodies to determine the features that correlate with the ability to neutralize toxin and to cooperate synergistically in toxin neutralization. The primary result of these experiments will be the creation of combinations of cloned human antibodies that can effectively counteract BoNT/A and BoNT/B exposure.