Xiong Xianghua, Qiu Yujin, Zheng Jiahao, Zhou Ling, Wang Qingyang, Pang Jinglun, Zhang Weicai, Chen Huipeng, Liu Gang, Han Xiaodong
Academy of Military Medical Sciences, Beijing, PR China.
Institute of Pharmaceutical Science, King's College London, 150 Stamford Street, London, SE1 9NH, UK.
Protein Expr Purif. 2024 Jan;213:106370. doi: 10.1016/j.pep.2023.106370. Epub 2023 Sep 13.
Botulinum neurotoxin serotype A (BoNT/A) can cause flaccid paralysis of muscles, an illness fatal to human, by entering neurons and blocking neurotransmitter release. The process was mediated by three receptors. A specific monoclonal antibody anti-D23, designated as ML419, targeting the ectodomain (D23) of fibroblast growth factor receptor 3 (FGFR3), one of the three receptors, was screened and capable of disturbing the recognition of BoNT/A and FGFR3. ML419 was screened from 14 stable positive hybridoma cell lines, and was subcloned, sequenced, and classified as IgG2a(κ) subclass. ML419 binds the D23 domain of FGFR3 with high affinity (K∼0.26 nM), and prevents the BoNT/A from entering Neuro-2a cells effectively. In vivo data showed that, 200 μg of ML419 could completely protect all the mice against with 5 MLD BoNT/A, while 100 μg of ML419 could protected 60% of the mice. Collectively, our results indicated that ML419 served as a good candidate for further development of therapeutics for BoNT/A.
A型肉毒杆菌神经毒素（BoNT/A）可通过进入神经元并阻断神经递质释放，导致肌肉弛缓性麻痹，这是一种对人类致命的疾病。该过程由三种受体介导。筛选出一种特异性抗D23单克隆抗体，命名为ML419，它靶向三种受体之一的成纤维细胞生长因子受体3（FGFR3）的胞外域（D23），并能够干扰BoNT/A与FGFR3的识别。ML419是从14个稳定的阳性杂交瘤细胞系中筛选出来的，经过亚克隆、测序，被归类为IgG2a(κ)亚类。ML419以高亲和力（K∼0.26 nM）结合FGFR3的D23结构域，并有效阻止BoNT/A进入Neuro-2a细胞。体内数据表明，200 μg的ML419可以完全保护所有小鼠免受5个最小致死量BoNT/A的侵害，而100 μg的ML419可以保护60%的小鼠。总的来说，我们的结果表明ML419是BoNT/A治疗药物进一步开发的良好候选物。
