| Autor: |
Bastos, Renata Chagas, Corrêa, Marilza Batista, de Souza, Iaralice Medeiros, da Silva, Milton Neto, da Silva Gomes Pereira, Denise, Martins, Fernanda Otaviano, da Silva Faria, Camila, Ano Bom, Ana Paula Dinis, de Lourdes Leal, Maria, Jessouroun, Ellen, da Silva, José Godinho, de Andrade Medronho, Ricardo, da Silveira, Ivna Alana Freitas Brasileiro |
| Zdroj: |
Glycoconjugate Journal; Feb2018, Vol. 35 Issue 1, p3-13, 11p |
| Abstrakt: |
High temperature is known to cause some instability in polysaccharide-protein conjugated vaccines and studies under stress conditions may be useful in determining whether short-term accidental exposure to undesired conditions can compromise product quality. In this study, we examined the structural stability of three industrial batches of Brazilian Meningococcal C conjugate bulk (MPCT) incubated at 4, 37, and 55 °C for 5 weeks. The effect of exposure to the storage temperatures was monitored by HPLC-SEC, CZE, CD and NMR techniques. The immunological significance of any physicochemical changes observed in MPCT was determined by SBA and ELISA assays of serum from immunized mice. Fluorescence emission spectra at 4 and 37 °C were similar among all samples and compatible with the native fold of the carrier protein. Fluorescence spectra of MPCT stored at 55 °C decreased in intensity and had a significant red-shift, indicating conformational changes. Far-UV CD spectra revealed a trend toward loss of structural conformation as storage temperature was increased to 55 °C. The NMR data showed modified signal intensity of the aromatic and aliphatic residues, mainly for samples incubated at 55 °C, suggesting a partial loss of tertiary structure. About 50% free saccharide content was found in bulks stored at 55 °C, but no difference was observed in the IgG or SBA titers. The present study showed physicochemical methods alone are insufficient to predict the biological activity of a MPCT conjugate vaccine without extensive validation against immunological data. However, they provide a sensitive means of detecting changes induced in a vaccine exposed to adverse environmental condition. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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