Production of Monoclonal Antibodies Against Botulinum Neurotoxin in

Overview

Journal Hum Vaccin Immunother

Specialty Allergy & Immunology

Date 2024 Mar 25

PMID 38525945

Authors

Kornchanok Sangprasat

Christine Joy I Bulaon

Kaewta Rattanapisit

Theerakarn Srisangsung

Perawat Jirarojwattana

Apidsada Wongwatanasin

Waranyoo Phoolcharoen

Affiliations

Soon will be listed here.

Abstract

Botulism is a fatal neurologic disease caused by the botulinum toxin (BoNT) produced by . 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 . 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.

Citing Articles

as a potential source for producing anti-dengue virus D54 neutralizing therapeutic antibody.

Krittanai S, Rattanapisit K, Bulaon C, Pitaksajjakul P, Keadsanti S, Ramasoota P Biotechnol Rep (Amst). 2024; 42:e00844.

PMID: 38881650 PMC: 11179242. DOI: 10.1016/j.btre.2024.e00844.

References

Zhang X, Han L, Zong H, Ding K, Yuan Y, Bai J . Enhanced production of anti-PD1 antibody in CHO cells through transient co-transfection with anti-apoptotic genes Bcl-x and Mcl-1. Bioprocess Biosyst Eng. 2018; 41(5):633-640. DOI: 10.1007/s00449-018-1898-z. View

Nowakowski A, Wang C, Powers D, Amersdorfer P, Smith T, Montgomery V . Potent neutralization of botulinum neurotoxin by recombinant oligoclonal antibody. Proc Natl Acad Sci U S A. 2002; 99(17):11346-50. PMC: 123259. DOI: 10.1073/pnas.172229899. View

Phakham T, Bulaon C, Khorattanakulchai N, Shanmugaraj B, Buranapraditkun S, Boonkrai C . Functional Characterization of Pembrolizumab Produced in Using a Rapid Transient Expression System. Front Plant Sci. 2021; 12:736299. PMC: 8459022. DOI: 10.3389/fpls.2021.736299. View

Helenius A, Aebi M . Intracellular functions of N-linked glycans. Science. 2001; 291(5512):2364-9. DOI: 10.1126/science.291.5512.2364. View

Arnon S, Schechter R, INGLESBY T, Henderson D, Bartlett J, Ascher M . Botulinum toxin as a biological weapon: medical and public health management. JAMA. 2001; 285(8):1059-70. DOI: 10.1001/jama.285.8.1059. View

Phetphoung T, Malla A, Rattanapisit K, Pisuttinusart N, Damrongyot N, Joyjamras K . Expression of plant-produced anti-PD-L1 antibody with anoikis sensitizing activity in human lung cancer cells via., suppression on epithelial-mesenchymal transition. PLoS One. 2022; 17(11):e0274737. PMC: 9651560. DOI: 10.1371/journal.pone.0274737. View

Chen Z, Morris Jr J, Rodriguez R, Wagle Shukla A, Tapia-Nunez J, Okun M . Emerging opportunities for serotypes of botulinum neurotoxins. Toxins (Basel). 2012; 4(11):1196-222. PMC: 3509704. DOI: 10.3390/toxins4111196. View

Kelley B . Industrialization of mAb production technology: the bioprocessing industry at a crossroads. MAbs. 2010; 1(5):443-52. PMC: 2759494. DOI: 10.4161/mabs.1.5.9448. View

Kroon C, Breuer L, Jones L, An J, Akan A, Ahmed Mohamed Ali E . Blind spots on western blots: Assessment of common problems in western blot figures and methods reporting with recommendations to improve them. PLoS Biol. 2022; 20(9):e3001783. PMC: 9518894. DOI: 10.1371/journal.pbio.3001783. View

10.

Tacket C, Shandera W, Mann J, Hargrett N, Blake P . Equine antitoxin use and other factors that predict outcome in type A foodborne botulism. Am J Med. 1984; 76(5):794-8. DOI: 10.1016/0002-9343(84)90988-4. View

11.

Rattanapisit K, Bulaon C, Strasser R, Sun H, Phoolcharoen W . In vitro and in vivo studies of plant-produced Atezolizumab as a potential immunotherapeutic antibody. Sci Rep. 2023; 13(1):14146. PMC: 10465495. DOI: 10.1038/s41598-023-41510-w. View

12.

Dressler D . Botulinum toxin mechanisms of action. Suppl Clin Neurophysiol. 2005; 57:159-66. DOI: 10.1016/s1567-424x(09)70353-8. View

13.

Matsumura T, Amatsu S, Misaki R, Yutani M, Du A, Kohda T . Fully Human Monoclonal Antibodies Effectively Neutralizing Botulinum Neurotoxin Serotype B. Toxins (Basel). 2020; 12(5). PMC: 7291131. DOI: 10.3390/toxins12050302. View

14.

Sack M, Rademacher T, Spiegel H, Boes A, Hellwig S, Drossard J . From gene to harvest: insights into upstream process development for the GMP production of a monoclonal antibody in transgenic tobacco plants. Plant Biotechnol J. 2015; 13(8):1094-105. DOI: 10.1111/pbi.12438. View

15.

Ahmadi S, Davami F, Davoudi N, Nematpour F, Ahmadi M, Ebadat S . Monoclonal antibodies expression improvement in CHO cells by PiggyBac transposition regarding vectors ratios and design. PLoS One. 2017; 12(6):e0179902. PMC: 5491063. DOI: 10.1371/journal.pone.0179902. View

16.

Saeidi S, Dadpour B, Jarahi L, Ghamsari A, Nooghabi M . Clinical Predictive Values in Botulism: A 10-year Survey. Indian J Crit Care Med. 2021; 25(4):411-415. PMC: 8138651. DOI: 10.5005/jp-journals-10071-23777. View

17.

Fox C, Keet C, Strober J . Recent advances in infant botulism. Pediatr Neurol. 2005; 32(3):149-54. DOI: 10.1016/j.pediatrneurol.2004.10.001. View

18.

Bulaon C, Khorattanakulchai N, Rattanapisit K, Sun H, Pisuttinusart N, Strasser R . Antitumor effect of plant-produced anti-CTLA-4 monoclonal antibody in a murine model of colon cancer. Front Plant Sci. 2023; 14:1149455. PMC: 10497774. DOI: 10.3389/fpls.2023.1149455. View

19.

Barash J, Arnon S . A novel strain of Clostridium botulinum that produces type B and type H botulinum toxins. J Infect Dis. 2013; 209(2):183-91. DOI: 10.1093/infdis/jit449. View

20.

Buyel J, Opdensteinen P, Fischer R . Cellulose-based filter aids increase the capacity of depth filters during the downstream processing of plant-derived biopharmaceutical proteins. Biotechnol J. 2015; 10(4):584-91. DOI: 10.1002/biot.201400611. View