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Ultra-Fast Middle-Up Reversed Phase Liquid Chromatography Analysis of Complex Bispecific Antibodies Obtained in Less Than One Minute
This work illustrates the benefits and limitations of using ultra-short reversed phase liquid chromatography (RPLC) columns for the characterization of various complex bispecific antibodies after prolonged thermal stress at the middle-up level of analysis. First, we have demonstrated that alternativ...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9698801/ https://www.ncbi.nlm.nih.gov/pubmed/36365134 http://dx.doi.org/10.3390/pharmaceutics14112315 |
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author | Murisier, Amarande D’Atri, Valentina Pirner, Sebastian Larraillet, Vincent Fekete, Szabolcs Lauber, Matthew Guillarme, Davy |
author_facet | Murisier, Amarande D’Atri, Valentina Pirner, Sebastian Larraillet, Vincent Fekete, Szabolcs Lauber, Matthew Guillarme, Davy |
author_sort | Murisier, Amarande |
collection | PubMed |
description | This work illustrates the benefits and limitations of using ultra-short reversed phase liquid chromatography (RPLC) columns for the characterization of various complex bispecific antibodies after prolonged thermal stress at the middle-up level of analysis. First, we have demonstrated that alternative organic modifiers, such as isopropanol, can be used in RPLC mode without generating excessive pressure, thanks to the prototype 10 × 2.1 mm, 2.7 µm particle column. However, compared to acetonitrile, the selectivity was not improved, at least for the selected biopharmaceutical products. Importantly, very fast separations (sub-1 min) of high quality were systematically obtained for the different samples when using a spectroscopic detector, but a severe loss of performance was observed with mass spectrometry (MS) detection due to dispersion effects. Based on these results, there is a clear need to improve the interfacing between LC and MS (shorter/thinner tubing) to mitigate band broadening. |
format | Online Article Text |
id | pubmed-9698801 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-96988012022-11-26 Ultra-Fast Middle-Up Reversed Phase Liquid Chromatography Analysis of Complex Bispecific Antibodies Obtained in Less Than One Minute Murisier, Amarande D’Atri, Valentina Pirner, Sebastian Larraillet, Vincent Fekete, Szabolcs Lauber, Matthew Guillarme, Davy Pharmaceutics Communication This work illustrates the benefits and limitations of using ultra-short reversed phase liquid chromatography (RPLC) columns for the characterization of various complex bispecific antibodies after prolonged thermal stress at the middle-up level of analysis. First, we have demonstrated that alternative organic modifiers, such as isopropanol, can be used in RPLC mode without generating excessive pressure, thanks to the prototype 10 × 2.1 mm, 2.7 µm particle column. However, compared to acetonitrile, the selectivity was not improved, at least for the selected biopharmaceutical products. Importantly, very fast separations (sub-1 min) of high quality were systematically obtained for the different samples when using a spectroscopic detector, but a severe loss of performance was observed with mass spectrometry (MS) detection due to dispersion effects. Based on these results, there is a clear need to improve the interfacing between LC and MS (shorter/thinner tubing) to mitigate band broadening. MDPI 2022-10-28 /pmc/articles/PMC9698801/ /pubmed/36365134 http://dx.doi.org/10.3390/pharmaceutics14112315 Text en © 2022 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 | Communication Murisier, Amarande D’Atri, Valentina Pirner, Sebastian Larraillet, Vincent Fekete, Szabolcs Lauber, Matthew Guillarme, Davy Ultra-Fast Middle-Up Reversed Phase Liquid Chromatography Analysis of Complex Bispecific Antibodies Obtained in Less Than One Minute |
title | Ultra-Fast Middle-Up Reversed Phase Liquid Chromatography Analysis of Complex Bispecific Antibodies Obtained in Less Than One Minute |
title_full | Ultra-Fast Middle-Up Reversed Phase Liquid Chromatography Analysis of Complex Bispecific Antibodies Obtained in Less Than One Minute |
title_fullStr | Ultra-Fast Middle-Up Reversed Phase Liquid Chromatography Analysis of Complex Bispecific Antibodies Obtained in Less Than One Minute |
title_full_unstemmed | Ultra-Fast Middle-Up Reversed Phase Liquid Chromatography Analysis of Complex Bispecific Antibodies Obtained in Less Than One Minute |
title_short | Ultra-Fast Middle-Up Reversed Phase Liquid Chromatography Analysis of Complex Bispecific Antibodies Obtained in Less Than One Minute |
title_sort | ultra-fast middle-up reversed phase liquid chromatography analysis of complex bispecific antibodies obtained in less than one minute |
topic | Communication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9698801/ https://www.ncbi.nlm.nih.gov/pubmed/36365134 http://dx.doi.org/10.3390/pharmaceutics14112315 |
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