Comparison and Optimization of Quantification Methods for <i>Shigella flexneri</i> Serotype 6 O-antigen Containing Galacturonic Acid and Methyl-Pentose
<i>Shigella</i> is a leading diarrheal cause of morbidity and mortality worldwide, especially in low- and middle-income countries and in children under five years of age. Increasing levels of antimicrobial resistance make vaccine development an even higher global health priority. <i&g...
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| Autores principales: | , , , |
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| Formato: | article |
| Lenguaje: | EN |
| Publicado: |
MDPI AG
2021
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| Materias: | |
| Acceso en línea: | https://doaj.org/article/0f77f58ebcc1413f9b61e7f2f82376bc |
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| Sumario: | <i>Shigella</i> is a leading diarrheal cause of morbidity and mortality worldwide, especially in low- and middle-income countries and in children under five years of age. Increasing levels of antimicrobial resistance make vaccine development an even higher global health priority. <i>S. flexneri</i> serotype 6 is one of the targets of many multicomponent vaccines in development to ensure broad protection against <i>Shigella</i>. The O-antigen (OAg) is a key active ingredient and its content is a critical quality attribute for vaccine release in order to monitor their stability and to ensure appropriate immune response. Here, the optimization of two methods to quantify <i>S. flexneri</i> 6 OAg is reported together with the characterization of their performances. The optimized Dische colorimetric method allows a tenfold increment of the sensitivity with respect to the original method and is useful for fast analysis detecting selectively methyl-pentoses, as rhamnose in <i>S. flexneri</i> 6 OAg. Also, a more specific HPAEC-PAD method was developed, detecting the dimer galacturonic acid-galactosamine (GalA-GalN) coming from <i>S. flexneri</i> 6 OAg acid hydrolysis. These methods will facilitate characterization of <i>S. flexneri</i> 6 OAg based vaccines. The colorimetric method can be used for quantification of other polysaccharide containing methyl-pentoses, and the HPAEC-PAD could be extended to other polysaccharides containing uronic acids. |
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