The European AntibotABE Framework Program and Its Update: Development of Innovative Botulinum Antibodies
Abstract
:1. Introduction
1.1. Background
1.2. Botulinum Neurotoxins (BoNTs)
1.3. Antitoxin Treatment
2. Animal Immunization and Antibody Phage-Display Library Construction
2.1. Immunization of Macaques (Macaca fascicularis) Using the Non-Toxic and Recombinant HC or LC of BoNT/A, B or E
2.2. Phage-Displayed Immune Libraries Construction
2.3. Selection of scFv Directed against BoNT/A1 and A2, or B1 and B2 or E3 Subtypes/Serotypes by Multi-Step Panning and Screening by ELISA
2.4. Further Characterization by Affinity Measurements
3. In Vitro Evaluations and Ex Vivo Neutralizing Properties of High Affinity scFvs and scFv-Fcs
3.1. In Vitro Screening of Anti-BoNT LC Antibodies as scFv or scFv-Fc by Endopeptidase Inhibition Assays
3.2. Ex Vivo Screening of Anti-BoNT HC and Anti-BoNT LC Antibodies as scFv or scFv-Fc in the Mouse Phrenic Nerve Hemidiaphragm Paralysis Assay
3.3. Isolation of the Most Promising Antibody Combinations (Anti-BoNT/HC and Anti-BoNT/LC Directed against the Same Serotype) Using the In Vivo Flaccid Paralysis Assay, Synergistic Protection Assessment, Epitope Characterization and Affinities Measurements
4. Germline-Humanization and In Vivo Characterization of the Selected Antibodies
4.1. Generation of Variants of the Germline Humanized Antibodies and Identification of the Most Promising Germline-Humanized Variant for Each Library
4.2. Expression of the Selected Germline-Humanized Variants as Full-Lengths IgGs
4.3. Assessment of the Protection Induced by Germline-Humanized IgGs in Lethal and Non-Lethal In Vivo Assays, Individually and in Combination
5. Output of the AntiBotABE Project
6. Dissemination of the AntiBotABE Project
- The first aim was to promote knowledge of our results among the potentials users of our antibodies, the governmental structures involved in bioterrorism preparedness, and the general public.
- In a next step, we addressed potential stakeholders for the further clinical and regulatory development of the AntiBotABE antibody cocktail (oligoclonal antibody). In order to draw the attention of European institutions, national governments, regional authorities and other public and private funding sources to the needs and benefits of our antibodies; and the need to stockpile them in advance: several presentations to scientific and decision-makers fora such as the European Defense Agency (EDA) and the North Atlantic Treaty Organization (NATO) have been made.
- Enhancing the reputation of participants at local, national and international levels and attracting the interest of correspondents, including the public; Aid the search for financial backers, licensees or industrial implementers to exploit the results and maintain market demand for the developed products or services. To this aim, regular up-dates of the AntiBotABE progress have been presented as oral presentations and posters at international meetings by the Consortium members.
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Antigen | Antibody | Format | In Vitro Inhibition: IC 50 (nM) | Ex Vivo Neutralization: Neutralizing Concentration 50% (nM) | Affinity (KD, nM) |
---|---|---|---|---|---|
BoNT/A LC | SEM120-IIIC1 | scFv-Fc | 10 | 1000 | 0.82 |
BoNT/A HC | AHC38 | scFv | n/a | 33 | 1.9 |
BoNT/B LC | BLC3 | scFv-Fc | 66 | 66 | 0.4 |
BoNT/B HC | B2-7 | scFv | n/a | >1000 | 4.8 |
BoNT/E LC | ELC18 | scFv | 112 | 3.3 | 0.58 |
BoNT/E HC | - | n/a | n/a | n/a | n/a |
Antigen | Antibody | Format | Germinality Index (GI %) |
---|---|---|---|
BoNT/A LC | SEM120-IIIC1 | scFv-Fc | 86.8 (VH)–87.6 (VL) |
BoNT/A HC | AHC38 | scFv | 86.5 (VH)–84.4 (VL) |
BoNT/B LC | BLC3 | scFv-Fc | 85.7 (VH)–85.7 (VL) |
BoNT/B HC | B2-7 | scFv | 85.7 (VH)–77.2 (VL) |
BoNT/E LC | ELC18 | scFv | 85.7 (VH)–89.9 (VL) |
BoNT/E HC | - | n/a | n/a |
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Rasetti-Escargueil, C.; Avril, A.; Miethe, S.; Mazuet, C.; Derman, Y.; Selby, K.; Thullier, P.; Pelat, T.; Urbain, R.; Fontayne, A.; et al. The European AntibotABE Framework Program and Its Update: Development of Innovative Botulinum Antibodies. Toxins 2017, 9, 309. https://doi.org/10.3390/toxins9100309
Rasetti-Escargueil C, Avril A, Miethe S, Mazuet C, Derman Y, Selby K, Thullier P, Pelat T, Urbain R, Fontayne A, et al. The European AntibotABE Framework Program and Its Update: Development of Innovative Botulinum Antibodies. Toxins. 2017; 9(10):309. https://doi.org/10.3390/toxins9100309
Chicago/Turabian StyleRasetti-Escargueil, Christine, Arnaud Avril, Sebastian Miethe, Christelle Mazuet, Yagmur Derman, Katja Selby, Philippe Thullier, Thibaut Pelat, Remi Urbain, Alexandre Fontayne, and et al. 2017. "The European AntibotABE Framework Program and Its Update: Development of Innovative Botulinum Antibodies" Toxins 9, no. 10: 309. https://doi.org/10.3390/toxins9100309
APA StyleRasetti-Escargueil, C., Avril, A., Miethe, S., Mazuet, C., Derman, Y., Selby, K., Thullier, P., Pelat, T., Urbain, R., Fontayne, A., Korkeala, H., Sesardic, D., Hust, M., & Popoff, M. R. (2017). The European AntibotABE Framework Program and Its Update: Development of Innovative Botulinum Antibodies. Toxins, 9(10), 309. https://doi.org/10.3390/toxins9100309