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http://hdl.handle.net/10773/41783
Title: | One step towards the quantification of p53-minicircle DNA |
Author: | Valente, Ana I. Tavares, Ana P. M. Sousa, Fani Freire, Mara G. |
Keywords: | Biopharmaceutical Minicircle DNA Extraction Quantification Ionic Liquids Citotoxicity |
Issue Date: | Jul-2023 |
Publisher: | UA Editora |
Abstract: | p53-minicircle DNA (mcDNA) is a biopharmaceutical with potential use in the genetic therapy of cancer. Despite the therapeutic relevance of DNA-based products, current clarification and purification steps are complex, resulting in high-cost biopharmaceuticals [1]. The existing strategies for mcDNA isolation are based on chromatographic techniques, such as size-exclusion, hydrophobic interaction, and affinity chromatography. Despite the selectivity achieved with the affinity strategy, it depends on the genetic manipulation of the vector to include specific sequences and/or in the use of enzymes to eliminate impurities [2,3]. Thus, up to date, one of the best strategies resorts to size exclusion chromatography, allowing the recovery of 66% of mcDNA with a purity of 98% [2]. Due to the lack of an efficient purification strategy, a quantification method for mcDNA is non-existent. Considering this, an analytical method based on ion exchange chromatography for mcDNA quantification is under development. Firstly, the p53-mcDNA is produced through the culture of transformed Escherichia coli. Then, the mcDNA is extracted with a commercial kit. The samples obtained after the extraction are applied in developing the new analytic method. Also, an extraction method using ionic liquid (IL)-based aqueous biphasic systems (ABS) is under evaluation for the replacement of the commercial kit used for the purpose. For that, the cytotoxicity of several analogues of glycine-betaine ILs is being also assessed. |
Peer review: | yes |
URI: | http://hdl.handle.net/10773/41783 |
ISBN: | 978-972-789-892-3 |
Publisher Version: | https://ria.ua.pt/handle/10773/39666 |
Appears in Collections: | CICECO - Comunicações DQ - Comunicações |
Files in This Item:
File | Description | Size | Format | |
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Valente et al_Research Summit_2023.pdf | 185.6 kB | Adobe PDF | View/Open |
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