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...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-10-01
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| Series: | Pharmaceutics |
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| Online Access: | https://www.mdpi.com/1999-4923/14/11/2315 |
| _version_ | 1797466837198307328 |
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| author | Amarande Murisier Valentina D’Atri Sebastian Pirner Vincent Larraillet Szabolcs Fekete Matthew Lauber Davy Guillarme |
| author_facet | Amarande Murisier Valentina D’Atri Sebastian Pirner Vincent Larraillet Szabolcs Fekete Matthew Lauber Davy Guillarme |
| author_sort | Amarande Murisier |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-09T18:45:20Z |
| format | Article |
| id | doaj.art-b56a7f4ec0ff4897b9a4111ffd99b447 |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-09T18:45:20Z |
| publishDate | 2022-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceutics |
| spelling | doaj.art-b56a7f4ec0ff4897b9a4111ffd99b4472023-11-24T06:20:12ZengMDPI AGPharmaceutics1999-49232022-10-011411231510.3390/pharmaceutics14112315Ultra-Fast Middle-Up Reversed Phase Liquid Chromatography Analysis of Complex Bispecific Antibodies Obtained in Less Than One MinuteAmarande Murisier0Valentina D’Atri1Sebastian Pirner2Vincent Larraillet3Szabolcs Fekete4Matthew Lauber5Davy Guillarme6School of Pharmaceutical Sciences, University of Geneva, CMU-Rue Michel Servet 1, 1211 Geneva, SwitzerlandSchool of Pharmaceutical Sciences, University of Geneva, CMU-Rue Michel Servet 1, 1211 Geneva, SwitzerlandRoche Diagnostics GmbH, Nonnenwald 2, 82377 Penzberg, GermanyRoche Diagnostics GmbH, Nonnenwald 2, 82377 Penzberg, GermanyWaters Corporation, CMU-Rue Michel Servet 1, 1211 Geneva, SwitzerlandWaters Corporation, 34 Maple Street, Milford, MA 01757-3696, USASchool of Pharmaceutical Sciences, University of Geneva, CMU-Rue Michel Servet 1, 1211 Geneva, SwitzerlandThis 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.https://www.mdpi.com/1999-4923/14/11/2315ultra-short columnsbispecific antibodiesreversed-phase liquid chromatographyimproved selectivitymethod robustness |
| spellingShingle | Amarande Murisier Valentina D’Atri Sebastian Pirner Vincent Larraillet Szabolcs Fekete Matthew Lauber Davy Guillarme Ultra-Fast Middle-Up Reversed Phase Liquid Chromatography Analysis of Complex Bispecific Antibodies Obtained in Less Than One Minute Pharmaceutics ultra-short columns bispecific antibodies reversed-phase liquid chromatography improved selectivity method robustness |
| 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 | ultra-short columns bispecific antibodies reversed-phase liquid chromatography improved selectivity method robustness |
| url | https://www.mdpi.com/1999-4923/14/11/2315 |
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