Production of monoclonal antibodies against botulinum neurotoxin in Nicotiana benthamiana

Autor: Kornchanok Sangprasat, Christine Joy I. Bulaon, Kaewta Rattanapisit, Theerakarn Srisangsung, Perawat Jirarojwattana, Apidsada Wongwatanasin, Waranyoo Phoolcharoen
Jazyk: angličtina
Rok vydání: 2024
Předmět:
Zdroj: Human Vaccines & Immunotherapeutics, Vol 20, Iss 1 (2024)
Druh dokumentu: article
ISSN: 21645515
2164-554X
2164-5515
55784925
DOI: 10.1080/21645515.2024.2329446
Popis: ABSTRACTBotulism is a fatal neurologic disease caused by the botulinum toxin (BoNT) produced by Clostridium botulinum. It is a rare but highly toxic disease with symptoms, such as cramps, nausea, vomiting, diarrhea, dysphagia, respiratory failure, muscle weakness, and even death. Currently, two types of antitoxin are used: equine-derived heptavalent antitoxin and human-derived immunoglobulin (BabyBIG®). However, heptavalent treatment may result in hypersensitivity, whereas BabyBIG®, has a low yield. The present study focused on the development of three anti-BoNT monoclonal antibodies (mAbs), 1B18, C25, and M2, in Nicotiana benthamiana. The plant-expressed mAbs were purified and examined for size, purity and integrity by SDS-PAGE, western blotting and size-exclusion chromatography. Analysis showed that plant-produced anti-BoNT mAbs can fully assemble in plants, can be purified in a single purification step, and mostly remain as monomeric proteins. The efficiency of anti-BoNT mAbs binding to BoNT/A and B was then tested. Plant-produced 1B18 retained its ability to recognize both mBoNT/A1 and ciBoNT/B1. At the same time, the binding specificities of two other mAbs were determined: C25 for mBoNT/A1 and M2 for ciBoNT/B1. In conclusion, our results confirm the use of plants as an alternative platform for the production of anti-BoNT mAbs. This plant-based technology will serve as a versatile system for the development botulism immunotherapeutics.
Databáze: Directory of Open Access Journals
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