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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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Autores principales: Carloni, Silvia, Piccinini, Claudia, Pancisi, Elena, Soldati, Valentina, Stefanelli, Monica, Granato, Anna Maria, Ibrahim, Toni, Petrini, Massimiliano
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8198694/
https://www.ncbi.nlm.nih.gov/pubmed/34072360
http://dx.doi.org/10.3390/ijms22115824
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author Carloni, Silvia
Piccinini, Claudia
Pancisi, Elena
Soldati, Valentina
Stefanelli, Monica
Granato, Anna Maria
Ibrahim, Toni
Petrini, Massimiliano
author_facet Carloni, Silvia
Piccinini, Claudia
Pancisi, Elena
Soldati, Valentina
Stefanelli, Monica
Granato, Anna Maria
Ibrahim, Toni
Petrini, Massimiliano
author_sort Carloni, Silvia
collection PubMed
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.
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spelling pubmed-81986942021-06-14 Potency Assessment of Dendritic Cell Anticancer Vaccine: Validation of the Co-Flow DC Assay Carloni, Silvia Piccinini, Claudia Pancisi, Elena Soldati, Valentina Stefanelli, Monica Granato, Anna Maria Ibrahim, Toni Petrini, Massimiliano Int J Mol Sci Article 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. MDPI 2021-05-29 /pmc/articles/PMC8198694/ /pubmed/34072360 http://dx.doi.org/10.3390/ijms22115824 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Carloni, Silvia
Piccinini, Claudia
Pancisi, Elena
Soldati, Valentina
Stefanelli, Monica
Granato, Anna Maria
Ibrahim, Toni
Petrini, Massimiliano
Potency Assessment of Dendritic Cell Anticancer Vaccine: Validation of the Co-Flow DC Assay
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 Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8198694/
https://www.ncbi.nlm.nih.gov/pubmed/34072360
http://dx.doi.org/10.3390/ijms22115824
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