Expression of an anti-botulinum toxin A neutralizing single-chain Fv recombinant antibody in transgenic tobacco

• Published in: Vaccine Vol. 24; no. 12; pp. 2079 - 2086
• Main Authors: Almquist, Kurt C., McLean, Michael D., Niu, Yongqing, Byrne, Greg, Olea-Popelka, Fernando C., Murrant, Coral, Barclay, Jack, Hall, J. Christopher
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Ltd 15.03.2006; Elsevier; Elsevier Limited
• Subjects: Advantages; Applied microbiology; Biological and medical sciences; Bioreactors; Botulinum Toxins, Type A - antagonists & inhibitors; Botulinum Toxins, Type A - genetics; Botulinum Toxins, Type A - immunology; Clearance; Cloning; Clostridium botulinum - chemistry; Clostridium botulinum - genetics; Clostridium botulinum - immunology; Costs; Deoxyribonucleic acid; DNA; Fundamental and applied biological sciences. Psychology; Genetic engineering; Immunoglobulin Fragments - genetics; Immunoglobulin Fragments - immunology; Immunoglobulin Fragments - metabolism; Microbiology; Neurotoxins; Nicotiana; Nicotiana tabacum; Pharmaceutical Preparations; Plants, Genetically Modified - genetics; Plants, Genetically Modified - metabolism; Proteins; Recombinant Fusion Proteins - immunology; Recombinant Fusion Proteins - metabolism; scFv; Tobacco; Toxin; Toxins; Transgenic plants; Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies (general aspects); Toxin; Tobacco; Clearance; scFv; Enzyme; Metalloendopeptidases; Stimulant plant; Nicotiana tabacum; Neutralizing antibody; Peptidases; Bontoxilysin; Dicotyledones; Angiospermae; Hydrolases; Spermatophyta; Solanaceae; Single chain antibody
• ISSN: 0264-410X; 1873-2518
• DOI: 10.1016/j.vaccine.2005.11.014

Botulinum neurotoxins (BoNTs) are the most poisonous substances known and are thus classified as high-risk threats for use as bioterror agents. To examine the potential of transgenic plants as bioreactors for the production of BoNT antidotes, we transformed tobacco with an optimized, synthetic gene encoding a botulinum neurotoxin A (BoNT/A) neutralizing single-chain Fv (scFv) recombinant antibody fragment. In vitro mouse muscle twitch assays demonstrated the functional utility of this scFv extracted from tobacco for neutralizing the paralytic effects of BoNT/A at neuromuscular junctions. Based on the efficiency of the scFv capture process and the dose required to antidote a human being, 1–2 ha of this tobacco could yield up to 4 kg of scFv, which would be enough to contribute to the manufacture of 1,000,000 therapeutic doses of a monoclonal antibody (mAb) cocktail capable of neutralizing the effects of BoNT poisoning. Transgenic plants could provide an inexpensive production platform for expression of multiple mAbs toward the creation of polyclonal therapies (i.e. pooled mAbs) as the next improvement in recombinant antibody therapy.