Potency Assessment of Dendritic Cell Anticancer Vaccine: Validation of the Co-Flow DC Assay
For many years, oncological clinical trials have taken advantage of dendritic cells (DC) for the design of DC-based cellular therapies. This has required the design of suitable quality control assays to evaluate the potency of these products. The purpose of our work was to develop and validate a nov...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-05-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/11/5824 |
| _version_ | 1797532114659311616 |
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| author | Silvia Carloni Claudia Piccinini Elena Pancisi Valentina Soldati Monica Stefanelli Anna Maria Granato Toni Ibrahim Massimiliano Petrini |
| author_facet | Silvia Carloni Claudia Piccinini Elena Pancisi Valentina Soldati Monica Stefanelli Anna Maria Granato Toni Ibrahim Massimiliano Petrini |
| author_sort | Silvia Carloni |
| collection | DOAJ |
| description | For many years, oncological clinical trials have taken advantage of dendritic cells (DC) for the design of DC-based cellular therapies. This has required the design of suitable quality control assays to evaluate the potency of these products. The purpose of our work was to develop and validate a novel bioassay that uses flow cytometry as a read-out measurement. In this method, CD3+ cells are labeled with a fluorescent dye and the DC costimulatory activity is measured by the degree of T cell proliferation caused by the DC–T cell interaction. The validation of the method was achieved by the evaluation of essential analytical parameters defined by international guidelines. Our results demonstrated that the method could be considered specific, selective, and robust. The comparison between measured values and estimated true values confirmed a high level of accuracy and a lack of systematic error. Repeated experiments have shown the reproducibility of the assay and the proportionality between the potency and the DC amount has proven its linearity. Our results suggest that the method is compliant with the guidelines and could be adopted as a quality control assay or batch-release testing within GMP facilities. |
| first_indexed | 2024-03-10T10:54:30Z |
| format | Article |
| id | doaj.art-479a6f98170342b9ad49cd58cdb7bfdf |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T10:54:30Z |
| publishDate | 2021-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-479a6f98170342b9ad49cd58cdb7bfdf2023-11-21T21:58:41ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-05-012211582410.3390/ijms22115824Potency Assessment of Dendritic Cell Anticancer Vaccine: Validation of the Co-Flow DC AssaySilvia Carloni0Claudia Piccinini1Elena Pancisi2Valentina Soldati3Monica Stefanelli4Anna Maria Granato5Toni Ibrahim6Massimiliano Petrini7Immuno-Gene Therapy Factory, IRCCS Istituto Romagnolo per lo Studio dei Tumori (IRST) “Dino Amadori”, 47014 Meldola, ItalyImmuno-Gene Therapy Factory, IRCCS Istituto Romagnolo per lo Studio dei Tumori (IRST) “Dino Amadori”, 47014 Meldola, ItalyImmuno-Gene Therapy Factory, IRCCS Istituto Romagnolo per lo Studio dei Tumori (IRST) “Dino Amadori”, 47014 Meldola, ItalyImmuno-Gene Therapy Factory, IRCCS Istituto Romagnolo per lo Studio dei Tumori (IRST) “Dino Amadori”, 47014 Meldola, ItalyImmuno-Gene Therapy Factory, IRCCS Istituto Romagnolo per lo Studio dei Tumori (IRST) “Dino Amadori”, 47014 Meldola, ItalyImmuno-Gene Therapy Factory, IRCCS Istituto Romagnolo per lo Studio dei Tumori (IRST) “Dino Amadori”, 47014 Meldola, ItalyOsteoncology and Rare Tumors Center, Immunotherapy, Cell Therapy and Biobank, IRCCS Istituto Romagnolo per lo Studio dei Tumori (IRST) “Dino Amadori”, 47014 Meldola, ItalyImmuno-Gene Therapy Factory, IRCCS Istituto Romagnolo per lo Studio dei Tumori (IRST) “Dino Amadori”, 47014 Meldola, ItalyFor many years, oncological clinical trials have taken advantage of dendritic cells (DC) for the design of DC-based cellular therapies. This has required the design of suitable quality control assays to evaluate the potency of these products. The purpose of our work was to develop and validate a novel bioassay that uses flow cytometry as a read-out measurement. In this method, CD3+ cells are labeled with a fluorescent dye and the DC costimulatory activity is measured by the degree of T cell proliferation caused by the DC–T cell interaction. The validation of the method was achieved by the evaluation of essential analytical parameters defined by international guidelines. Our results demonstrated that the method could be considered specific, selective, and robust. The comparison between measured values and estimated true values confirmed a high level of accuracy and a lack of systematic error. Repeated experiments have shown the reproducibility of the assay and the proportionality between the potency and the DC amount has proven its linearity. Our results suggest that the method is compliant with the guidelines and could be adopted as a quality control assay or batch-release testing within GMP facilities.https://www.mdpi.com/1422-0067/22/11/5824dendritic cellspotencyT cell proliferationvalidationco-stimulationflow cytometry |
| spellingShingle | Silvia Carloni Claudia Piccinini Elena Pancisi Valentina Soldati Monica Stefanelli Anna Maria Granato Toni Ibrahim Massimiliano Petrini Potency Assessment of Dendritic Cell Anticancer Vaccine: Validation of the Co-Flow DC Assay International Journal of Molecular Sciences dendritic cells potency T cell proliferation validation co-stimulation flow cytometry |
| title | Potency Assessment of Dendritic Cell Anticancer Vaccine: Validation of the Co-Flow DC Assay |
| title_full | Potency Assessment of Dendritic Cell Anticancer Vaccine: Validation of the Co-Flow DC Assay |
| title_fullStr | Potency Assessment of Dendritic Cell Anticancer Vaccine: Validation of the Co-Flow DC Assay |
| title_full_unstemmed | Potency Assessment of Dendritic Cell Anticancer Vaccine: Validation of the Co-Flow DC Assay |
| title_short | Potency Assessment of Dendritic Cell Anticancer Vaccine: Validation of the Co-Flow DC Assay |
| title_sort | potency assessment of dendritic cell anticancer vaccine validation of the co flow dc assay |
| topic | dendritic cells potency T cell proliferation validation co-stimulation flow cytometry |
| url | https://www.mdpi.com/1422-0067/22/11/5824 |
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