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Review
. 2017 Oct 2;9(10):309.
doi: 10.3390/toxins9100309.

The European AntibotABE Framework Program and Its Update: Development of Innovative Botulinum Antibodies

Christine Rasetti-Escargueil  1 , Arnaud Avril  2   3 , Sebastian Miethe  4 , Christelle Mazuet  5 , Yagmur Derman  6 , Katja Selby  7 , Philippe Thullier  8 , Thibaut Pelat  9   10 , Remi Urbain  11   12 , Alexandre Fontayne  13 , Hannu Korkeala  14 , Dorothea Sesardic  15 , Michael Hust  16 , Michel R Popoff  17
Affiliations
Review

The European AntibotABE Framework Program and Its Update: Development of Innovative Botulinum Antibodies

Christine Rasetti-Escargueil et al. Toxins (Basel). .

Abstract

The goal of the AntiBotABE Program was the development of recombinant antibodies that neutralize botulinum neurotoxins (BoNT) A, B and E. These serotypes are lethal and responsible for most human botulinum cases. To improve therapeutic efficacy, the heavy and light chains (HC and LC) of the three BoNT serotypes were targeted to achieve a synergistic effect (oligoclonal antibodies). For antibody isolation, macaques were immunized with the recombinant and non-toxic BoNT/A, B or E, HC or LC, followed by the generation of immune phage-display libraries. Antibodies were selected from these libraries against the holotoxin and further analyzed in in vitro and ex vivo assays. For each library, the best ex vivo neutralizing antibody fragments were germline-humanized and expressed as immunoglobulin G (IgGs). The IgGs were tested in vivo, in a standardized model of protection, and challenged with toxins obtained from collections of Clostridium strains. Protective antibody combinations against BoNT/A and BoNT/B were evidenced and for BoNT/E, the anti-LC antibody alone was found highly protective. The combination of these five antibodies as an oligoclonal antibody cocktail can be clinically and regulatorily developed while their high "humanness" predicts a high tolerance in humans.

Keywords: AntiBotABE; IgG; biodefense; botulinum; botulism; neutralization; oligoclonal antibodies; phage-display; recombinant; toxin.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
AntiBotABE Workflow.
Figure 2
Figure 2
ELISA on immobilized BoNT/A LC. 1 μg of the 19 selected binders against BoNT/A1 tested as scFv-Fc fusion (murine Fc) on 100 ng directly immobilized BoNT/A LC. Modified figure from our previous publication: Miethe et al. 2014 [51].
Figure 3
Figure 3
Neutralization of BoNT/A1 by scFv-Fc directed against BoNT/A LC on the mouse phrenic nerve-hemidiaphragm. Modified figure from our previous publication: Miethe et al. 2014 [51].
Figure 4
Figure 4
Neutralization activity in MPNH of scFv-Fc B2-7 targeting BoNT/B-HC. Modified figure from our previous publication: Rasetti-Escargueil et al. 2015 [74].
Figure 5
Figure 5
Neutralization activity of scFv-Fc BLC3 and B2-7 in vivo. Modified figure from our previous publication: Rasetti-Escargueil et al. 2015 [74].
Figure 6
Figure 6
In vivo mouse paralysis assay with ELC18 as scFv-Fc. Modified figure from our previous publication: Miethe et al. 2015 [75].
Figure 7
Figure 7
Antigen binding of humanized variants of SEM120-IIIC1 (anti-BoNT/A1 LC).
Figure 8
Figure 8
ELISA assay of all humanized ELC18 variants (from hu1ELC18 to hu8ELC18) and non-humanized ELC18. Modified figure from our previous publication: Derman et al. 2016 [76].
Figure 9
Figure 9
Survival rate of 5 MLD50/mice with a range of antibody concentrations.
Figure 10
Figure 10
Overview of AntiBotABE output.
See this image and copyright information in PMC

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