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Agglomeration behaviour of magnetic microparticles during separation and recycling processes in mRNA purification
Purification of mRNA with oligo(dT)‐functionalized magnetic particles involves a series of magnetic separations for buffer exchange and washing. Magnetic particles interact and agglomerate with each other when a magnetic field is applied, which can result in a decreased total surface area and thus a...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8518558/ https://www.ncbi.nlm.nih.gov/pubmed/34690629 http://dx.doi.org/10.1002/elsc.202000112 |
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author | Wommer, Lars Soerjawinata, Winda Ulber, Roland Kampeis, Percy |
author_facet | Wommer, Lars Soerjawinata, Winda Ulber, Roland Kampeis, Percy |
author_sort | Wommer, Lars |
collection | PubMed |
description | Purification of mRNA with oligo(dT)‐functionalized magnetic particles involves a series of magnetic separations for buffer exchange and washing. Magnetic particles interact and agglomerate with each other when a magnetic field is applied, which can result in a decreased total surface area and thus a decreased yield of mRNA. In addition, agglomeration may also be caused by mRNA loading on the magnetic particles. Therefore, it is of interest how the individual steps of magnetic separation and subsequent redispersion in the buffers used affect the particle size distribution. The lysis/binding buffer is the most important buffer for the separation of mRNA from the multicomponent suspension of cell lysate. Therefore, monodisperse magnetic particles loaded with mRNA were dispersed in the lysis/binding buffer and in the reference system deionized water, and the particle size distributions were measured. A concentration‐dependent agglomeration tendency was observed in deionized water. In contrast, no significant agglomeration was detected in the lysis/binding buffer. With regard to magnetic particle recycling, the influence of different storage and drying processes on particle size distribution was investigated. Agglomeration occurred in all process alternatives. For de‐agglomeration, ultrasonic treatment was examined. It represents a suitable method for reproducible restoration of the original particle size distribution. |
format | Online Article Text |
id | pubmed-8518558 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-85185582021-10-22 Agglomeration behaviour of magnetic microparticles during separation and recycling processes in mRNA purification Wommer, Lars Soerjawinata, Winda Ulber, Roland Kampeis, Percy Eng Life Sci Research Articles Purification of mRNA with oligo(dT)‐functionalized magnetic particles involves a series of magnetic separations for buffer exchange and washing. Magnetic particles interact and agglomerate with each other when a magnetic field is applied, which can result in a decreased total surface area and thus a decreased yield of mRNA. In addition, agglomeration may also be caused by mRNA loading on the magnetic particles. Therefore, it is of interest how the individual steps of magnetic separation and subsequent redispersion in the buffers used affect the particle size distribution. The lysis/binding buffer is the most important buffer for the separation of mRNA from the multicomponent suspension of cell lysate. Therefore, monodisperse magnetic particles loaded with mRNA were dispersed in the lysis/binding buffer and in the reference system deionized water, and the particle size distributions were measured. A concentration‐dependent agglomeration tendency was observed in deionized water. In contrast, no significant agglomeration was detected in the lysis/binding buffer. With regard to magnetic particle recycling, the influence of different storage and drying processes on particle size distribution was investigated. Agglomeration occurred in all process alternatives. For de‐agglomeration, ultrasonic treatment was examined. It represents a suitable method for reproducible restoration of the original particle size distribution. John Wiley and Sons Inc. 2021-06-24 /pmc/articles/PMC8518558/ /pubmed/34690629 http://dx.doi.org/10.1002/elsc.202000112 Text en © 2021 The Authors. Engineering in Life Sciences published by Wiley‐VCH GmbH https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Articles Wommer, Lars Soerjawinata, Winda Ulber, Roland Kampeis, Percy Agglomeration behaviour of magnetic microparticles during separation and recycling processes in mRNA purification |
title | Agglomeration behaviour of magnetic microparticles during separation and recycling processes in mRNA purification |
title_full | Agglomeration behaviour of magnetic microparticles during separation and recycling processes in mRNA purification |
title_fullStr | Agglomeration behaviour of magnetic microparticles during separation and recycling processes in mRNA purification |
title_full_unstemmed | Agglomeration behaviour of magnetic microparticles during separation and recycling processes in mRNA purification |
title_short | Agglomeration behaviour of magnetic microparticles during separation and recycling processes in mRNA purification |
title_sort | agglomeration behaviour of magnetic microparticles during separation and recycling processes in mrna purification |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8518558/ https://www.ncbi.nlm.nih.gov/pubmed/34690629 http://dx.doi.org/10.1002/elsc.202000112 |
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