New approach to generating of human monoclonal antibodies specific to the proteolytic domain of botulinum neurotoxin A

• Published in: Bioimpacts 2024, Vol.14 (4), p.1-14
• Main Authors: Silkina, Marina Vladimirovna, Kartseva, Alena Sergeevna, Riabko, Alena Konstantinovna, Makarova, Mariia Aleksandrovna, Rogozin, Metkhun Madibronovich, Romanenko, Yana Olegovna, Shemyakin, Igor Georgievich, Dyatlov, Ivan Alekseevich, Firstova, Victoria Valerievna
• Format: Article
• Language: English
• Subjects: Acetylcholine; Anaphylaxis; Antisera; Botulinum toxin type A; Botulism; Children; Cranial nerves; Flow cytometry; Ganglia; Immunotherapy; Molecular weight; Monoclonal antibodies; Multiple myeloma; Nerve endings; Neuromuscular junctions; Original; Paralysis; Peripheral nerves; Polypeptides; Proteins; Proteolysis; Public health; Toxins; Ventilation
• ISSN: 2228-5652; 2228-5660
• DOI: 10.34172/bi.2023.27680

BoNT enters the vascular circulation and is transported to peripheral nerve terminals, neuromuscular junctions and peripheral ganglia.6 As a result of blocking the transmission of acetylcholine through the neuromuscular junction by inhibiting the release of acetylcholine from the presynaptic end of the motor neuron, paralysis occurs.7 Through similar pharmacological mechanisms at the neuromuscular junction share and a similar structure all BoNT types produce a similar clinical syndrome of cranial nerve palsies that can be followed by bilateral flaccid paralysis, affecting limb musculature, that might progress to respiratory failure and death.3,7,8 The BoNT molecule is a heterodimer synthesised as a single polypeptide chain with a molecular weight of 150 kDa, consisting of a 100 kDa heavy chain (HC) and a 50 kDa light chain (LC) linked by a disulfide bond.9 HC consists of two functionally distinct regions: the receptor-binding C-terminal domain (HC50) ~50 kDa and the translocation N-terminal domain (HN). Bering botulism antitoxin (Novartis Vaccines and Diagnostics GmbH and Co. KG) is used to treat types A, B and E, while heptavalent botulism antitoxin (BAT, Cangene Corporation) is used to treat types A, B, C, D, E, F and G. However, as concentrated preparations of foreign proteins, they can trigger immune reactions, including anaphylaxis.15,16 A formulation of a human immune serum called BabyBIG (California Department of Public Health) has recently become available, but in limited supply and for treating only childhood botulism.17,18 In addition, immune serum production involves complex and time-consuming production processes and quality management. [...]the development of safe, effective, and standardized antibodies is required. The heated culture (1 mL) was inoculated into the toxin production broth medium containing 30 g/L Trypton T (Type III, HiMedia, India), 5 g/L glucose, 20 g/L yeast extract and 0.5 g/L L-cysteine HCl, pH 7.4. [...]the precipitate was redissolved in 0.05 M citrate-phosphate buffer with pH 5.5.38 The resulting native BoNT/A was used for Western blot and FRET assay.