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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...

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Autores principales: Wommer, Lars, Soerjawinata, Winda, Ulber, Roland, Kampeis, Percy
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
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.
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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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