Development of a Monocyte Activation Test as an Alternative to the Rabbit Pyrogen Test for Mono- and Multi-Component Shigella GMMA-Based Vaccines
Open Access
- 24 June 2021
- journal article
- research article
- Published by MDPI AG in Microorganisms
- Vol. 9 (7), 1375
- https://doi.org/10.3390/microorganisms9071375
Abstract
Generalised modules for membrane antigens (GMMA)-based vaccines comprise the outer membrane from genetically modified Gram-negative bacteria containing membrane proteins, phospholipids and lipopolysaccharides. Some lipoproteins and lipopolysaccharides are pyrogens; thus, GMMA-based vaccines are intrinsically pyrogenic. It is important to control the pyrogenic content of biological medicines, including vaccines, to prevent adverse reactions such as febrile responses. The rabbit pyrogen test (RPT) and bacterial endotoxin test (BET) are the most commonly employed safety assays used to detect pyrogens. However, both tests are tailored for detecting pyrogenic contaminants and have considerable limitations when measuring the pyrogen content of inherently pyrogenic products. We report the adaptation of the monocyte activation test (MAT) as an alternative to the RPT for monitoring the pyrogenicity of Shigella GMMA-based vaccines. The European Pharmacopoeia endorses three MAT methods (A–C). Of these, method C, the reference lot comparison test, was identified as the most suitable. This method was evaluated with different reference materials to ensure parallelism and consistency for a mono- and multi-component Shigella GMMA vaccine. We demonstrate the drug substance as a promising reference material for safety testing of the matched drug product. Our results support the implementation of MAT as an alternative to the RPT and use of the defined parameters can be extended to GMMA-based vaccines currently in development, aiding vaccine batch release.Keywords
Funding Information
- UK Department of Health and Social Care (EP/R013764/1)
This publication has 32 references indexed in Scilit:
- An improved monocyte activation test using cryopreserved pooled human mononuclear cellsInnate Immunity, 2015
- The human whole blood pyrogen test – lessons learned in twenty yearsAltex, 2015
- Modulation of Endotoxicity of Shigella Generalized Modules for Membrane Antigens (GMMA) by Genetic Lipid A ModificationsOnline Journal of Public Health Informatics, 2014
- A Comparative Review of Toll-Like Receptor 4 Expression and Functionality in Different Animal SpeciesFrontiers in Immunology, 2014
- High Yield Production Process for Shigella Outer Membrane ParticlesPLOS ONE, 2012
- Improved in vitro methods to predict the in vivo toxicity in man of therapeutic monoclonal antibodies including TGN1412Journal of Immunological Methods, 2010
- International validation of novel pyrogen tests based on human monocytoid cellsJournal of Immunological Methods, 2005
- Effects of aluminum adjuvant on systemic reactions of lipopolysaccharides in swineVaccine, 1995
- DETECTION OF PYROGEN BY CYTOKINE RELEASEThe Lancet, 1988
- The detection of endotoxin by in vitro production of endogenous pyrogen: Comparison with limulus amebocyte lysate gelationJournal of Immunological Methods, 1982