Design and evaluation of mRNA encoding recombinant neutralizing antibodies for botulinum neurotoxin type B intoxication prophylaxis

ABSTRACT

Among all natural and synthetic toxins, botulinum neurotoxins (BoNTs), produced by Clostridium botulinum in an anaerobic environment, are the most toxic polymer proteins. Currently, the most effective modalities for botulism prevention and treatment are vaccination and antitoxin use, respectively. However, these modalities are associated with long response time for active immunization, side effects, and donor limitations. As such, the development of more promising botulism prevention and treatment modalities is warranted. Here, we designed an mRNA encoding B9-hFc – a heavy-chain antibody fused to VHH and human Fc that can neutralize BoNT serotype B (BoNT/B) effectively – and assessed its expression in vitro and in vivo. The results confirmed that our mRNA demonstrates good expression in vitro and in vivo. Moreover, a single mRNA lipid nanoparticle injection effectively prevents BoNT/B intoxication in vivo, with effects comparable to those of protein antibodies. In conclusion, we explored and clarified whether mRNA drugs encoding neutralizing antibodies prevent BoNT/B intoxication. Our results provide an efficient strategy for further research on the prevention and treatment of intoxication by botulinum toxin.

KEYWORDS:

• Botulinum neurotoxin
• mRNA
• prophylaxis
• heavy-chain antibody
• lipid nanoparticle

Acknowledgments

We thank Dr. Xiaofan Zhao (Beijing Institute of Biotechnology) for support of the LNPs coating technique.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Author contributions

Conceptualization, P.D., J.L and G.Q.; methodology, G.Q., Y.J. and G.L.; software, G.Q.; formal analysis, G.Q., P.D. and J.L.; investigation, G.Q. and Y.J.; resources, G.Q. and G.L.; data curation, G.Q.; writ-ing – original draft preparation, G.Q., Y.J. and J.L.; writing – review and editing, Y.C., P.D., G.Q., J.L., Z.Y., F.L. and Y.Y.; supervision, P.D., Y.C., J.L., Z.Y.and F.L.; project administration, Y.C., P.D., J.L. and G.Q. All authors have read and agreed to the published version of the manuscript.

Data availability statement

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

Supplementary material

Supplemental data for this article can be accessed on the publisher’s website at https://doi.org/10.1080/21645515.2024.2358570

Additional information

Funding

The author(s) reported there is no funding associated with the work featured in this article.