Method for the Quantification of Alginate in Microcapsules
Alginate is a key reagent in the preparation of microcapsules for cell transplantation. To address the question of the intracapsular alginate concentration, a sensitive assay has been developed to quantify the alginate content of microcapsules. The method is based on the metachromatic change induced...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
SAGE Publishing
1993-09-01
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| Series: | Cell Transplantation |
| Online Access: | https://doi.org/10.1177/096368979300200511 |
| _version_ | 1818146596900044800 |
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| author | Jean-Pierre Hallé Dr. Danielle Landry Alain Fournier Michèle Beaudry Francois A. Leblond |
| author_facet | Jean-Pierre Hallé Dr. Danielle Landry Alain Fournier Michèle Beaudry Francois A. Leblond |
| author_sort | Jean-Pierre Hallé Dr. |
| collection | DOAJ |
| description | Alginate is a key reagent in the preparation of microcapsules for cell transplantation. To address the question of the intracapsular alginate concentration, a sensitive assay has been developed to quantify the alginate content of microcapsules. The method is based on the metachromatic change induced by alginate binding to the dye, 1,9-dimethyl methylene blue (DMMB). The assay has a high sensitivity and precision. It covers a wide concentration range enabling the measurement of alginate in dilute supernatants as well as in microcapsules. For the latter, the membrane is initially dissolved by incubating the microcapsules in an alkaline medium. The effect of potentially interfering substances (poly-l-lysine (PLL), citrate, chloride, sodium) and of pH has been studied. Poly-l-lysine interfered with the assay at pH 6.5 but not at pH 13. Interference by sodium augmented with increasing sodium concentration and reached a plateau at 200 mM. This problem was overcome by routinely adjusting all samples to 500 mM sodium. The other substances tested had a negligible effect on the assay. The reliable measurement of alginate with this new assay will allow the optimization of the intracapsular alginate concentration. |
| first_indexed | 2024-12-11T12:21:52Z |
| format | Article |
| id | doaj.art-ec715e6f912549999bc57762d6dc490d |
| institution | Directory Open Access Journal |
| issn | 0963-6897 1555-3892 |
| language | English |
| last_indexed | 2024-12-11T12:21:52Z |
| publishDate | 1993-09-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Cell Transplantation |
| spelling | doaj.art-ec715e6f912549999bc57762d6dc490d2022-12-22T01:07:30ZengSAGE PublishingCell Transplantation0963-68971555-38921993-09-01210.1177/096368979300200511Method for the Quantification of Alginate in MicrocapsulesJean-Pierre Hallé Dr.0Danielle Landry1Alain Fournier2Michèle Beaudry3Francois A. Leblond4Centre de Recherche de L'Hôpital Maisonneuve-Rosemont, 5415, Boul. de l'Assomption, Montreal, Quebec, Canada, H1T 2M4Centre de Recherche de L'Hôpital Maisonneuve-Rosemont, 5415, Boul. de l'Assomption, Montreal, Quebec, Canada, H1T 2M4Institut National de la Recherche Scientifique-Santé 245 Boul. Hymus, Pointe-Claire, Quebec, Canada, H9R 1G6Centre de Recherche de L'Hôpital Maisonneuve-Rosemont, 5415, Boul. de l'Assomption, Montreal, Quebec, Canada, H1T 2M4Centre de Recherche de L'Hôpital Maisonneuve-Rosemont, 5415, Boul. de l'Assomption, Montreal, Quebec, Canada, H1T 2M4Alginate is a key reagent in the preparation of microcapsules for cell transplantation. To address the question of the intracapsular alginate concentration, a sensitive assay has been developed to quantify the alginate content of microcapsules. The method is based on the metachromatic change induced by alginate binding to the dye, 1,9-dimethyl methylene blue (DMMB). The assay has a high sensitivity and precision. It covers a wide concentration range enabling the measurement of alginate in dilute supernatants as well as in microcapsules. For the latter, the membrane is initially dissolved by incubating the microcapsules in an alkaline medium. The effect of potentially interfering substances (poly-l-lysine (PLL), citrate, chloride, sodium) and of pH has been studied. Poly-l-lysine interfered with the assay at pH 6.5 but not at pH 13. Interference by sodium augmented with increasing sodium concentration and reached a plateau at 200 mM. This problem was overcome by routinely adjusting all samples to 500 mM sodium. The other substances tested had a negligible effect on the assay. The reliable measurement of alginate with this new assay will allow the optimization of the intracapsular alginate concentration.https://doi.org/10.1177/096368979300200511 |
| spellingShingle | Jean-Pierre Hallé Dr. Danielle Landry Alain Fournier Michèle Beaudry Francois A. Leblond Method for the Quantification of Alginate in Microcapsules Cell Transplantation |
| title | Method for the Quantification of Alginate in Microcapsules |
| title_full | Method for the Quantification of Alginate in Microcapsules |
| title_fullStr | Method for the Quantification of Alginate in Microcapsules |
| title_full_unstemmed | Method for the Quantification of Alginate in Microcapsules |
| title_short | Method for the Quantification of Alginate in Microcapsules |
| title_sort | method for the quantification of alginate in microcapsules |
| url | https://doi.org/10.1177/096368979300200511 |
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