Structural organization of NadADelta(351-405), a recombinant MenB vaccine component, by its physico-chemical characterization at drug substance level

The physico-chemical characterization of NadADelta(351-405), a recombinant protein discovered by reverse vaccinology, component of a candidate vaccine against Neisseria meningitidis serotype B is presented. Analytical methods like mass spectrometry, electrophoresis, optical spectroscopy and SEC-MALL...

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Bibliographic Details
Published in:Vaccine 2009-03, Vol.27 (15), p.2156-2170
Main Authors: Magagnoli, Claudia, Bardotti, Angela, De Conciliis, Giuseppe, Galasso, Rosy, Tomei, Matteo, Campa, Cristiana, Pennatini, Carlo, Cerchioni, Maruska, Fabbri, Barbara, Giannini, Sara, Mattioli, Giovanni L, Biolchi, Alessia, D'Ascenzi, Sandro, Helling, Friedhelm
Format: Article
Language:English
Subjects:
Adhesins, Bacterial - chemistry
Adhesins, Bacterial - immunology
Amino Acid Sequence
Animals
Circular Dichroism
Female
Meningococcal Vaccines - chemistry
Meningococcal Vaccines - immunology
Mice
Molecular Sequence Data
Neisseria meningitidis, Serogroup B - chemistry
Neisseria meningitidis, Serogroup B - immunology
Peptide Mapping
Protein Conformation
Recombinant Proteins - chemistry
Recombinant Proteins - immunology
Spectrometry, Fluorescence
Vaccines, Synthetic - chemistry
Vaccines, Synthetic - immunology
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Summary:The physico-chemical characterization of NadADelta(351-405), a recombinant protein discovered by reverse vaccinology, component of a candidate vaccine against Neisseria meningitidis serotype B is presented. Analytical methods like mass spectrometry, electrophoresis, optical spectroscopy and SEC-MALLS have been applied to unveil the structure of NadADelta(351-405), and to evaluate Product-Related Substances. Moreover, analysis of the protein after intentional denaturation has been applied in order to challenge the chosen methods and to determine their appropriateness and specificity. All the obtained results were inserted in a model allowing in-depth understanding of the antigen NadADelta(351-405): it is present in solution as a homo-trimer, retaining a high percentage of alpha-helix secondary structure, and able to reassemble from monomeric subunits after thermal denaturation; this structural organization is consistent with that foreseen for MenB NadA (Neisseria Adhesin A). The analytical data sets produced during process development for clinical phases I-III material confirm product quality and manufacturing consistency.
ISSN:0264-410X
DOI:10.1016/j.vaccine.2009.01.099