Tehran University of Medical Sciences

Science Communicator Platform

Stay connected! Follow us on X network (Twitter):
Share this content! On (X network) By
Production and Characterization of Pertussis Toxin Specific Monoclonal and Polyclonal Antibodies: Implication for Toxin Purification and Detection Publisher Pubmed



Imani D1 ; Bahadori T1 ; Mobini M1 ; Judaki MA1 ; Jedditehrani M2 ; Amiri MM1 ; Shokri F1
Authors
Show Affiliations
Authors Affiliations
  1. 1. Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
  2. 2. Monoclonal Antibody Research Center, Avicenna Research Institute, Academic Center for Education, Culture and Research (ACECR), Tehran, Iran

Source: Toxicon Published:2025


Abstract

Background: Pertussis is a pulmonary disease caused by the gram-negative bacteria Bordetella pertussis (BP) with a high fatality rate among newborns and young children. Pertussis toxin (PT) is essential for pertussis pathogenesis as well as production of acellular pertussis vaccines (aPV). Traditional PT purification procedures are laborious and yield low purity and recovery rates. Also, due to the low production levels of PT by BP and the difficulties of purification, an appropriate immunoassay is needed to monitor PT concentrations upstream and downstream of the production process. This study investigates production and application of monoclonal and polyclonal antibodies for efficient PT purification and quantification. Methods: Rabbits and mice were immunized with native PT to produce polyclonal and monoclonal antibodies (MAbs). The MAbs were selected based on affinity, isotype and specificity, as determined by enzyme-linked immunosorbent assay (ELISA) and immunoblotting. The native PT antigen was purified using an immunoaffinity column. The purity and recovery rates of native PT were analyzed by ELISA, SDS-PAGE, and immunoblotting. Additionally, monoclonal and polyclonal antibodies were used to establish an ELISA assay for measurement of PT concentration. Results: A highly pure PT with recovery rates of around 74 ± 4.9 % was obtained following purification by immunoaffinity column, using polyclonal antibodies. Furthermore, the designed ELISA demonstrated suitable reactivity for measurement of the PT antigen. Conclusion: Our results indicate suitability of the produced monoclonal and polyclonal anti-PT antibodies for purification and monitoring of PT by immunoaffinity chromatography and ELISA, respectively. The immunoaffinity method offers an efficient replacement for PT purification in the context of developing aPV. © 2025
Related Docs
1. High Purity and Recovery of Native Filamentous Hemagglutinin (Fha) From Bordetella Pertussis Using Affinity Chromatography, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences (2024)
2. Novel Mouse Monoclonal Antibodies Against Bordetella Pertussis Pertactin Antigen With Versatile Applications, Journal of Microbiological Methods (2023)
Experts (# of related papers)
View all Related Experts
Masoumeh Douraghi (2)
Forough Golsaz Shirazi (2)