Analytical Insights into Protein–Alum Interactions and Their Impact on Conformational Epitope
Several alum-adjuvanted vaccines have been licensed in the past 40 years. Despite its extensive and continuous use, the immune mechanism of action of alum adjuvants is not yet completely understood. Many different variables during the formulation process have been assessed as critical for alum-adjuv...
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| Format: | Article |
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MDPI AG
2024-03-01
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| Series: | Pharmaceutics |
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| Online Access: | https://www.mdpi.com/1999-4923/16/3/420 |
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| author | Alessio Corrado Mila Toppazzini Alessandro Vadi Carmine Malzone Rosy Galasso Alessandro Donati Riccardo De Ricco Francesco Berti |
| author_facet | Alessio Corrado Mila Toppazzini Alessandro Vadi Carmine Malzone Rosy Galasso Alessandro Donati Riccardo De Ricco Francesco Berti |
| author_sort | Alessio Corrado |
| collection | DOAJ |
| description | Several alum-adjuvanted vaccines have been licensed in the past 40 years. Despite its extensive and continuous use, the immune mechanism of action of alum adjuvants is not yet completely understood. Many different variables during the formulation process have been assessed as critical for alum-adjuvanted vaccines, although most of them are still not yet fully understood. The absence of a clear understanding of all the possible variables regulating the mechanism of action and the behavior that alum adjuvant imposes on the protein antigen may also be related to analytical challenges. For this reason, there is an urgent need for a fast and simple tool that is possible without a preliminary sample manipulation and is able to control the amount and the degree of antigen adsorption levels and their consistency across different production processes. This work attempts to develop new analytical tools with the aim of directly quantifying and assessing both the content and/or the purity of formulated alum-adsorbed antigens, without any preliminary sample manipulation (e.g., antigen desorption) being reported. In addition, the different confirmation/behavior in terms of the response to specific monoclonal antibodies in the presence of different ratios of alum-OH adsorbent antigens have been investigated. As a proxy to develop new analytical tools, three recombinant protein adsorbed models were used as follows: Neisseria adhesin A (NadA), Neisserial Heparin Binding Antigen (NHBA), and factor H binding protein (fHbp) as antigens, as well as aluminum hydroxide (AH) as an adjuvant system. The selection of the adjuvanted system model was dictated due to the substantial quantity of the literature regarding the protein structure and immunological activities, meaning that they are well characterized, including their adhesion rate to alum. In conclusion, three different analytical tools were explored to quantify, detect, and study the behavior of antigens in the presence of the alum adjuvant. |
| first_indexed | 2024-04-24T17:54:48Z |
| format | Article |
| id | doaj.art-3282d5331ade4825bcdb3a34c887a3f1 |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-04-24T17:54:48Z |
| publishDate | 2024-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceutics |
| spelling | doaj.art-3282d5331ade4825bcdb3a34c887a3f12024-03-27T13:59:52ZengMDPI AGPharmaceutics1999-49232024-03-0116342010.3390/pharmaceutics16030420Analytical Insights into Protein–Alum Interactions and Their Impact on Conformational EpitopeAlessio Corrado0Mila Toppazzini1Alessandro Vadi2Carmine Malzone3Rosy Galasso4Alessandro Donati5Riccardo De Ricco6Francesco Berti7GSK, Vaccines Srl, via Fiorentina 1, 53100 Siena, ItalyGSK, Vaccines Srl, via Fiorentina 1, 53100 Siena, ItalyGSK, Vaccines Srl, via Fiorentina 1, 53100 Siena, ItalyGSK, Vaccines Srl, via Fiorentina 1, 53100 Siena, ItalyGSK, Vaccines Srl, via Fiorentina 1, 53100 Siena, ItalyDepartment of Biotechnology, Chemistry and Pharmacy, University of Siena, via A. Moro 2, 53100 Siena, ItalyGSK, Vaccines Srl, via Fiorentina 1, 53100 Siena, ItalyGSK, Vaccines Srl, via Fiorentina 1, 53100 Siena, ItalySeveral alum-adjuvanted vaccines have been licensed in the past 40 years. Despite its extensive and continuous use, the immune mechanism of action of alum adjuvants is not yet completely understood. Many different variables during the formulation process have been assessed as critical for alum-adjuvanted vaccines, although most of them are still not yet fully understood. The absence of a clear understanding of all the possible variables regulating the mechanism of action and the behavior that alum adjuvant imposes on the protein antigen may also be related to analytical challenges. For this reason, there is an urgent need for a fast and simple tool that is possible without a preliminary sample manipulation and is able to control the amount and the degree of antigen adsorption levels and their consistency across different production processes. This work attempts to develop new analytical tools with the aim of directly quantifying and assessing both the content and/or the purity of formulated alum-adsorbed antigens, without any preliminary sample manipulation (e.g., antigen desorption) being reported. In addition, the different confirmation/behavior in terms of the response to specific monoclonal antibodies in the presence of different ratios of alum-OH adsorbent antigens have been investigated. As a proxy to develop new analytical tools, three recombinant protein adsorbed models were used as follows: Neisseria adhesin A (NadA), Neisserial Heparin Binding Antigen (NHBA), and factor H binding protein (fHbp) as antigens, as well as aluminum hydroxide (AH) as an adjuvant system. The selection of the adjuvanted system model was dictated due to the substantial quantity of the literature regarding the protein structure and immunological activities, meaning that they are well characterized, including their adhesion rate to alum. In conclusion, three different analytical tools were explored to quantify, detect, and study the behavior of antigens in the presence of the alum adjuvant.https://www.mdpi.com/1999-4923/16/3/420bacterial vaccinesadjuvantprotein subunitvesiclesmeningococcal |
| spellingShingle | Alessio Corrado Mila Toppazzini Alessandro Vadi Carmine Malzone Rosy Galasso Alessandro Donati Riccardo De Ricco Francesco Berti Analytical Insights into Protein–Alum Interactions and Their Impact on Conformational Epitope Pharmaceutics bacterial vaccines adjuvant protein subunit vesicles meningococcal |
| title | Analytical Insights into Protein–Alum Interactions and Their Impact on Conformational Epitope |
| title_full | Analytical Insights into Protein–Alum Interactions and Their Impact on Conformational Epitope |
| title_fullStr | Analytical Insights into Protein–Alum Interactions and Their Impact on Conformational Epitope |
| title_full_unstemmed | Analytical Insights into Protein–Alum Interactions and Their Impact on Conformational Epitope |
| title_short | Analytical Insights into Protein–Alum Interactions and Their Impact on Conformational Epitope |
| title_sort | analytical insights into protein alum interactions and their impact on conformational epitope |
| topic | bacterial vaccines adjuvant protein subunit vesicles meningococcal |
| url | https://www.mdpi.com/1999-4923/16/3/420 |
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