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-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.
Novartis Vaccines, Via Fiorentina 1, 53100 Siena, Italy. claudia.magagnoli@novartis.com
Claudia
C
Magagnoli
Angela
A
Bardotti
Giuseppe
G
De Conciliis
Rosy
R
Galasso
Matteo
M
Tomei
Cristiana
C
Campa
Carlo
C
Pennatini
Maruska
M
Cerchioni
Barbara
B
Fabbri
Sara
S
Giannini
Giovanni L
GL
Mattioli
Alessia
A
Biolchi
Sandro
S
D'Ascenzi
Friedhelm
F
Helling
D018829:Adhesins, Bacterial / Q000737:chemistry* / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D002942:Circular Dichroism; D005260:Female; D022401:Meningococcal Vaccines / Q000737:chemistry* / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D038541:Neisseria meningitidis, Serogroup B / Q000737:chemistry* / Q000276:immunology; D010449:Peptide Mapping; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry* / Q000276:immunology; D013050:Spectrometry, Fluorescence; D014614:Vaccines, Synthetic / Q000737:chemistry / Q000276:immunology
D018829:Adhesins, Bacterial; D022401:Meningococcal Vaccines; C501982:NadA protein, Neisseria meningitidis; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic