Development of a New Solid-Phase Extraction Base Method for Free Saccharide Content Estimation of Meningococcal Conjugate Vaccines

Ricco, Riccardo De; Rech, Francesca; Onnis, Valeria; Coccone, Salvatore Sanna; Scalia, Giusy; Marcozzi, Cristina; Gavini, Massimiliano; Beni, Sara M.; Giannini, Sara; Nompari, Luca; Parlati, Chiara; Magagnoli, Claudia; Cianetti, Simona

doi:10.1021/acsomega.2c04013

Cited by 3 publications

(5 citation statements)

References 22 publications

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“…Micro BCA (Thermo Scientific, Waltham, MA, USA) and HPAEC-PAD [ 35 , 36 , 37 ] were used to estimate total protein and total PS content, respectively, and to calculate the saccharide-to-protein ratios. Free PS in GAC glycoconjugates was quantified by HPAEC-PAD after separation through DOC precipitation [ 38 ], while free Vi was separated by Capto Adhere resin and quantified by HPAEC-PAD [ 36 ].…”

Section: Methodsmentioning

confidence: 99%

“…Free PS in GAC glycoconjugates was quantified by HPAEC-PAD after separation through DOC precipitation [ 38 ], while free Vi was separated by Capto Adhere resin and quantified by HPAEC-PAD [ 36 ]. Free OSs in Men glycoconjugates were separated by solid phase extraction (SPE) and quantified by HPAEC-PAD [ 37 ]. Glycoconjugates formation was verified by HPLC-SEC and SDS-PAGE analyses, comparing the glycoconjugates with corresponding unconjugated proteins.…”

Section: Methodsmentioning

confidence: 99%

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Testing a Recombinant Form of Tetanus Toxoid as a Carrier Protein for Glycoconjugate Vaccines

Oldrini,

Di Benedetto,

Carducci

et al. 2023

Vaccines

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Glycoconjugate vaccines play a major role in the prevention of infectious diseases worldwide, with significant impact on global health, enabling the polysaccharides to induce immunogenicity in infants and immunological memory. Tetanus toxoid (TT), a chemically detoxified bacterial toxin, is among the few carrier proteins used in licensed glycoconjugate vaccines. The recombinant full-length 8MTT was engineered in E. coli with eight individual amino acid mutations to inactivate three toxin functions. Previous studies in mice showed that 8MTT elicits a strong IgG response, confers protection, and can be used as a carrier protein. Here, we compared 8MTT to traditional carrier proteins TT and cross-reactive material 197 (CRM197), using different polysaccharides as models: Group A Streptococcus cell-wall carbohydrate (GAC), Salmonella Typhi Vi, and Neisseria meningitidis serogroups A, C, W, and Y. The persistency of the antibodies induced, the ability of the glycoconjugates to elicit booster response after re-injection at a later time point, the eventual carrier-induced epitopic suppression, and immune interference in multicomponent formulations were also evaluated. Overall, immunogenicity responses obtained with 8MTT glycoconjugates were compared to those obtained with corresponding TT and, in some cases, were higher than those induced by CRM197 glycoconjugates. Our results support the use of 8MTT as a good alternative carrier protein for glycoconjugate vaccines, with advantages in terms of manufacturability compared to TT.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Testing a Recombinant Form of Tetanus Toxoid as a Carrier Protein for Glycoconjugate Vaccines

Oldrini,

Di Benedetto,

Carducci

et al. 2023

Vaccines

Self Cite

View full text Add to dashboard Cite

show abstract

“…In addition, a hydrolysis step is usually polysaccharide specific as it depends on the repeating unit structure, properties, and glycosidic linkage. It is also not possible to identify a single monosaccharide which is conserved between all polysaccharide antigens (neither across the Spn serotypes), leading to serotype specific hydrolysis and chromatography (as already observed for N. meningitidis serogroups A, C, W and Y 12 , 14 , 15 ).…”

Section: Introductionmentioning

confidence: 99%

“…In addition, the analysis of underivatized monosaccharides is becoming more and more relevant. For this purpose, high performance anion exchange chromatography with pulsed amphoteric detection (HPAEC-PAD) has become a reliable tool for the direct analysis of monosaccharides (routinely used by worldwide Official Medicines Control Laboratories OMCLs) 10 , 11 . HPAEC-PAD is characterized by very high levels of precision, reproducibility, and specificity for saccharide quantification.…”

Section: Introductionmentioning

confidence: 99%

“…On the other hand, long polysaccharide antigens are difficult to analyze without any major sample pre-treatment steps to either (i) purify the saccharide from impurities / matrix (e.g. Solid Phase Extraction treatment) and to (ii) cleave the glycosidic bonds to release the monosaccharide units 12 , 13 . In addition, a hydrolysis step is usually polysaccharide specific as it depends on the repeating unit structure, properties, and glycosidic linkage.…”

Section: Introductionmentioning

confidence: 99%

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A universal UHPLC-CAD platform for the quantification of polysaccharide antigens

Corrado

Martino

Bordoni

et al. 2023

Sci Rep

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Several glycoconjugate-based vaccines against bacterial infections have been developed and licensed for human use. Polysaccharide (PS) analysis and characterization is therefore critical to profile the composition of polysaccharide-based vaccines. For PS content quantification, the majority of Ultra High Performance Liquid Chromatography (UHPLC) methods rely on the detection of selective monosaccharides constituting the PS repeating unit, therefore requiring chemical cleavage and tailored development: only a few methods directly quantify the intact PS. The introduction of charged aerosol detector (CAD) technology has improved the response of polysaccharide analytes, offering greater sensitivity than other detector sources (e.g., ELSD). Herein, we report the development of a universal UHPLC-CAD method (UniQS) for the quantification and quality evaluation of polysaccharide antigens (e.g., Streptococcus Pneumoniae, Neisseria meningitidis and Staphylococcus aureus). This work laid the foundation for a universal UHPLC-CAD format that could play an important role in future vaccine research and development helping to reduce time, efforts, and costs.

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Method development and validation of unbound saccharide content (serogroup A, C, Y, W, X) in novel pentavalent meningococcal polysaccharide conjugate vaccine with two different carrier proteins

Kale,

Phugare,

Sharma

et al. 2023

Journal of Pharmaceutical and Biomedical Analysis

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Development of a New Solid-Phase Extraction Base Method for Free Saccharide Content Estimation of Meningococcal Conjugate Vaccines

Cited by 3 publications

References 22 publications

Testing a Recombinant Form of Tetanus Toxoid as a Carrier Protein for Glycoconjugate Vaccines

Testing a Recombinant Form of Tetanus Toxoid as a Carrier Protein for Glycoconjugate Vaccines

A universal UHPLC-CAD platform for the quantification of polysaccharide antigens

Method development and validation of unbound saccharide content (serogroup A, C, Y, W, X) in novel pentavalent meningococcal polysaccharide conjugate vaccine with two different carrier proteins

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