| Autor: |
Michiels TJM; Leiden Academic Centre for Drug Research (LACDR), Division of BioTherapeutics, Leiden University, 2333 CC Leiden, The Netherlands.; Intravacc, Institute for Translational Vaccinology, 3721 MA Bilthoven, The Netherlands., Tilstra W; Intravacc, Institute for Translational Vaccinology, 3721 MA Bilthoven, The Netherlands., Hamzink MRJ; Intravacc, Institute for Translational Vaccinology, 3721 MA Bilthoven, The Netherlands., de Ridder JW; Intravacc, Institute for Translational Vaccinology, 3721 MA Bilthoven, The Netherlands., Danial M; Intravacc, Institute for Translational Vaccinology, 3721 MA Bilthoven, The Netherlands., Meiring HD; Intravacc, Institute for Translational Vaccinology, 3721 MA Bilthoven, The Netherlands., Kersten GFA; Leiden Academic Centre for Drug Research (LACDR), Division of BioTherapeutics, Leiden University, 2333 CC Leiden, The Netherlands.; Intravacc, Institute for Translational Vaccinology, 3721 MA Bilthoven, The Netherlands., Jiskoot W; Leiden Academic Centre for Drug Research (LACDR), Division of BioTherapeutics, Leiden University, 2333 CC Leiden, The Netherlands., Metz B; Intravacc, Institute for Translational Vaccinology, 3721 MA Bilthoven, The Netherlands. |
| Abstrakt: |
Currently, batch release of toxoid vaccines, such as diphtheria and tetanus toxoid, requires animal tests to confirm safety and immunogenicity. Efforts are being made to replace these tests with in vitro assays in a consistency approach. Limitations of current in vitro assays include the need for reference antigens and most are only applicable to drug substance, not to the aluminum adjuvant-containing and often multivalent drug product. To overcome these issues, a new assay was developed based on mimicking the proteolytic degradation processes in antigen-presenting cells with recombinant cathepsin S, followed by absolute quantification of the formed peptides by liquid chromatography-mass spectrometry. Temperature-exposed tetanus toxoids from several manufacturers were used as aberrant samples and could easily be distinguished from the untreated controls by using the newly developed degradomics assay. Consistency of various batches of a single manufacturer could also be determined. Moreover, the assay was shown to be applicable to Al(OH) 3 and AlPO 4 -adsorbed tetanus toxoids. Overall, the assay shows potential for use in both stability studies and as an alternative for in vivo potency studies by showing batch-to-batch consistency of bulk toxoids as well as for aluminum-containing vaccines. |
