Single-domain antibody delivery using an mRNA platform protects against lethal doses of botulinum neurotoxin A

Panova, E. A.; Kleymenov, D. A.; Shcheblyakov, D. V.; Bykonia, E. N.; Mazunina, E. P.; Dzharullaeva, A. S.; Zolotar, A. N.; Derkaev, A. A.; Esmagambetov, I. B.; Sorokin, I. I.; Usachev, E. V.; Ivanov, I. A.; Zatsepin, T. S.; Dmitriev, S. E.; Gushchin, V. A.; Logunov, D. Y.; Gintsburg, A. L.

doi:10.1101/2022.10.29.514343

bioRxiv · 10.1101/2022.10.29.514343

Single-domain antibody delivery using an mRNA platform protects against lethal doses of botulinum neurotoxin A

Panova, E. A.·Kleymenov, D. A.·Shcheblyakov, D. V.·Bykonia, E. N.·Mazunina, E. P.·Dzharullaeva, A. S.·Zolotar, A. N.·Derkaev, A. A.·Esmagambetov, I. B.·Sorokin, I. I.·Usachev, E. V.·Ivanov, I. A.·Zatsepin, T. S.·Dmitriev, S. E.·Gushchin, V. A.·Logunov, D. Y.·Gintsburg, A. L.

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Abstract

Single-domain antibodies (sdAbs, VHHs, or nanobodies) are a promising tool for the treatment of both infectious and somatic diseases. Their small size greatly simplifies any genetic engineering manipulations. Such antibodies have the ability to bind hard-to-reach antigenic epitopes through long parts of the variable chains, the third complementarity-determining regions (CDR3s). VHH fusion with the canonical immunoglobulin Fc fragment allows the Fc-fusion single-domain antibodies (VHH-Fc) to significantly increase their neutralizing activity and serum half-life. Previously we have developed and characterized VHH-Fc specific to botulinum neurotoxin A (BoNT/A), that showed a 1000-fold higher protective activity than monomeric form when challenged with five times the lethal dose (5 LD50) of BoNT/A. During the COVID-19 pandemic, mRNA vaccines based on lipid nanoparticles (LNP) as a delivery system have become an important translational technology that has significantly accelerated the clinical introduction of mRNA platforms. We have developed an mRNA platform that provides long-term expression after both intramuscular and intravenous application. The platform has been extensively characterized using firefly luciferase (Fluc) as a reporter. An intramuscular administration of LNP-mRNA encoding VHH-Fc antibody made it possible to achieve its rapid expression in mice and resulted in 100% protection when challenged with up to 100 LD50 of BoNT/A. The presented approach for the delivery of sdAbs using mRNA technology greatly simplifies drug development for antibody therapy and can be used for emergency prophylaxis.

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Panova, E. A., Kleymenov, D. A., Shcheblyakov, D. V., Bykonia, E. N., Mazunina, E. P., Dzharullaeva, A. S., Zolotar, A. N., Derkaev, A. A., Esmagambetov, I. B., Sorokin, I. I., Usachev, E. V., Ivanov, I. A., Zatsepin, T. S., Dmitriev, S. E., Gushchin, V. A., Logunov, D. Y., Gintsburg, A. L.. 2022-10-30. Single-domain antibody delivery using an mRNA platform protects against lethal doses of botulinum neurotoxin A. https://doi.org/10.1101/2022.10.29.514343

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