Generation and characterization of a monoclonal antibody against FGFR3 that protects mice from BoNT/A

Xianghua Xiong; Yujin Qiu; Jiahao Zheng; Ling Zhou; Qingyang Wang; Jinglun Pang; Weicai Zhang; Huipeng Chen; Gang Liu; Xiaodong Han

doi:10.1016/j.pep.2023.106370

Generation and characterization of a monoclonal antibody against FGFR3 that protects mice from BoNT/A

Protein Expr Purif. 2024 Jan:213:106370. doi: 10.1016/j.pep.2023.106370. Epub 2023 Sep 13.

Authors

Xianghua Xiong¹, Yujin Qiu¹, Jiahao Zheng², Ling Zhou³, Qingyang Wang¹, Jinglun Pang⁴, Weicai Zhang¹, Huipeng Chen⁵, Gang Liu⁶, Xiaodong Han⁷

Affiliations

¹ Academy of Military Medical Sciences, Beijing, PR China.
² Institute of Pharmaceutical Science, King's College London, 150 Stamford Street, London, SE1 9NH, UK.
³ Department of Clinical Laboratory, The Affiliated Pudong Hospital, Fudan University, Shanghai, China.
⁴ College of Life Sciences, Inner Mongolia Agriculture University, Hohhot, Inner Mongolia, PR China.
⁵ Academy of Military Medical Sciences, Beijing, PR China. Electronic address: huipengchen@yeah.net.
⁶ Academy of Military Medical Sciences, Beijing, PR China. Electronic address: jueliu@sohu.com.
⁷ College of Life Sciences, Inner Mongolia Agriculture University, Hohhot, Inner Mongolia, PR China. Electronic address: hxdon@imau.edu.cn.

PMID: 37709211
DOI: 10.1016/j.pep.2023.106370

Abstract

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_D∼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 MLD₅₀ 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.

Keywords: Binding affinity; Botulinum neurotoxin A; Fibroblast growth factor receptor 3; Monoclonal antibody; Mortality rate.