Bivalent recombinant vaccine for botulinum neurotoxin types A and B based on a polypeptide comprising their effector and translocation domains that is protective against the predominant A and B subtypes

Clifford Shone, Heidi Agostini, Joanna Clancy, Mili Gu, Huei Hsiung Yang, Yanfang Chu, Virginia Johnson, Makie Taal, Joanna McGlashan, John Brehm, Xiaomi Tong*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

19 Citations (Scopus)

Abstract

The botulinum neurotoxins (BoNTs) are a large family of extremely potent, neuroparalytic, dichain proteins which act at the peripheral nervous system. The wide genetic diversity observed with this neurotoxin family poses a significant challenge for the development of an effective botulinum vaccine. The present study describes a vaccine development platform based on protein fragments representing the N-terminal two-thirds of each toxin molecule. These fragments, designated LHN, comprise the light chain and translocation domains of each neurotoxin and are devoid of any neuron-binding activity. Using codon-optimized genes, LHN fragments derived from BoNT serotypes A and B were expressed in Escherichia coli in high yield with >1 g of purified, soluble fragment recoverable from 4.5 liter-scale fermentations. The protective efficacy of LHN/A was significantly enhanced by treatment with formaldehyde, which induced intramolecular cross-linking but virtually no aggregation of the fragment. A single immunization of the modified fragment protected mice from challenge with a 103 50% lethal dose (LD 50) of BoNT/A1 with an 50% effective dose (ED 50) of 50 ng of the vaccine. In similar experiments, the LH N/A vaccine was shown to protect mice against challenge with BoNT/A subtypes A1, A2, and A3, which is the first demonstration of single-dose protection by a vaccine against the principal toxin subtypes of BoNT/A. The LHN/B vaccine was also highly efficacious, giving an ED50 of ∼140 ng to a challenge of 103 LD50 of BoNT/B1. In addition, LHN/B provided single-dose protection in mice against BoNT/B4 (non-proteolytic toxin subtype).

Original languageEnglish
Pages (from-to)2795-2801
Number of pages7
JournalInfection and Immunity
Volume77
Issue number7
DOIs
Publication statusPublished - Jul 2009

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