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:	Maria Michelina Raso, Oscar Vassallo, Francesca Micoli, Carlo Giannelli
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	<i>S. flexneri</i> O-antigen HPAEC-PAD Dische colorimetric method quantification polysaccharide vaccine Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/0f77f58ebcc1413f9b61e7f2f82376bc
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spelling	oai:doaj.org-article:0f77f58ebcc1413f9b61e7f2f82376bc2021-11-25T17:53:54ZComparison and Optimization of Quantification Methods for <i>Shigella flexneri</i> Serotype 6 O-antigen Containing Galacturonic Acid and Methyl-Pentose10.3390/ijms2222121601422-00671661-6596https://doaj.org/article/0f77f58ebcc1413f9b61e7f2f82376bc2021-11-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/22/12160https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067<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.Maria Michelina RasoOscar VassalloFrancesca MicoliCarlo GiannelliMDPI AGarticle<i>S. flexneri</i> O-antigenHPAEC-PADDische colorimetric methodquantificationpolysaccharidevaccineBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 12160, p 12160 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	<i>S. flexneri</i> O-antigen HPAEC-PAD Dische colorimetric method quantification polysaccharide vaccine Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	<i>S. flexneri</i> O-antigen HPAEC-PAD Dische colorimetric method quantification polysaccharide vaccine Biology (General) QH301-705.5 Chemistry QD1-999 Maria Michelina Raso Oscar Vassallo Francesca Micoli Carlo Giannelli Comparison and Optimization of Quantification Methods for <i>Shigella flexneri</i> Serotype 6 O-antigen Containing Galacturonic Acid and Methyl-Pentose
description	<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.
format	article
author	Maria Michelina Raso Oscar Vassallo Francesca Micoli Carlo Giannelli
author_facet	Maria Michelina Raso Oscar Vassallo Francesca Micoli Carlo Giannelli
author_sort	Maria Michelina Raso
title	Comparison and Optimization of Quantification Methods for <i>Shigella flexneri</i> Serotype 6 O-antigen Containing Galacturonic Acid and Methyl-Pentose
title_short	Comparison and Optimization of Quantification Methods for <i>Shigella flexneri</i> Serotype 6 O-antigen Containing Galacturonic Acid and Methyl-Pentose
title_full	Comparison and Optimization of Quantification Methods for <i>Shigella flexneri</i> Serotype 6 O-antigen Containing Galacturonic Acid and Methyl-Pentose
title_fullStr	Comparison and Optimization of Quantification Methods for <i>Shigella flexneri</i> Serotype 6 O-antigen Containing Galacturonic Acid and Methyl-Pentose
title_full_unstemmed	Comparison and Optimization of Quantification Methods for <i>Shigella flexneri</i> Serotype 6 O-antigen Containing Galacturonic Acid and Methyl-Pentose
title_sort	comparison and optimization of quantification methods for <i>shigella flexneri</i> serotype 6 o-antigen containing galacturonic acid and methyl-pentose
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/0f77f58ebcc1413f9b61e7f2f82376bc
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Comparison and Optimization of Quantification Methods for <i>Shigella flexneri</i> Serotype 6 O-antigen Containing Galacturonic Acid and Methyl-Pentose

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