In Vivo Neutralization of Botulinum Neurotoxins Serotype E with Heavy-chain Camelid Antibodies (VHH)
In Vivo Neutralization of Botulinum Neurotoxins Serotype E with Heavy-chain Camelid Antibodies (VHH)
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DOI：
10.1007/s12033-013-9669-1
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发表时间：
2013-10-01
影响因子：
2.6
通讯作者：
Zare, Hamed
中科院分区：
文献类型：
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作者：
Bakherad, Hamid;Gargari, Seyed Latif Mousavi;Zare, Hamed
肉毒杆菌神经毒素（BoNT）的摄入导致肉毒中毒，肉毒中毒是世界上已知的严重且频繁的致命疾病。目前的治疗依赖于抗毒素，如马抗毒素和人肉毒杆菌免疫球蛋白。在某些情况下，副作用已被报道，包括早期过敏性休克和晚期血清病。因此，BoNT的诊断和治疗措施是必要的和关键的。在本研究中，一个单域可变重链（VHH）抗体片段获得从免疫单峰骆驼噬菌体展示库对肉毒杆菌神经毒素E（BoNT/E），一个无毒的50 kDa片段的推定结合域。纳米抗体VHH的特性包括优异的生产、上级热稳定性和对可溶性抗原的特异性结合能力，而不与其他相关或不相关抗原发生交叉反应。在小鼠的体内激发中，总共150 ng/Kg的纳米抗体完全中和了3LD 50的BoNT/E。这种现象表明所产生的纳米抗体的BoNT/E毒素中和能力。这些结果还表明纳米抗体具有独特的性质，可用于诊断或治疗目的。
Ingestion of botulinum neurotoxin (BoNT) results in botulism, a severe and frequent fatal disease known in the world. Current treatments rely on antitoxins, such as equine antitoxin and human botulism immunoglobulin. In some cases, side effects have been reported, including early anaphylactic shock and late serum sickness. Thus, diagnosis and treatment measure of BoNT are necessary and crucial. In the present study, a single-domain variable heavy-chain (VHH) antibody fragment was obtained from an immune dromedary phage display library against the putative binding domain of botulinum neurotoxin E (BoNT/E), a non-toxic 50-kDa fragment. The characteristics of nanobody VHH include excellent production, superior heat stability and specific binding capacity to soluble antigen without cross-reaction to other relevant or irrelevant antigens. A total of 150 ng/Kg of nanobody entirely neutralized 3LD50 of the BoNT/E in an in vivo challenge of the mice. This phenomenon indicates BoNT/E toxin neutralizing capacity of the produced nanobody. These results also suggest possession of unique properties by the nanobody applicable in diagnostics or therapeutic purposes.