Interlaced size exclusion liquid chromatography of monoclonal antibodies
| Autor: | Guillaume Tremintin, Dell Farnan, George Tony Moreno, John T. Stults, Mark van Gils, Andreas Becker |
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| Rok vydání: | 2009 |
| Předmět: |
Time Factors
Chromatography Resolution (mass spectrometry) Elution medicine.drug_class Chemistry Sample (material) Organic Chemistry Size-exclusion chromatography Antibodies Monoclonal Reproducibility of Results General Medicine Monoclonal antibody Biochemistry Column (database) Analytical Chemistry Robustness (computer science) Chromatography Gel medicine Throughput (business) Chromatography Liquid |
| Zdroj: | Journal of Chromatography A. 1216:8904-8909 |
| ISSN: | 0021-9673 |
| Popis: | Size exclusion chromatography is a widely performed analysis of monoclonal antibodies, primarily used to monitor the levels of higher weight molecular species such as aggregates. Owing to the subtleties of these separation mechanisms and frequently observed partial resolutions of components in these separations, many common methods for increasing the method throughput are not practical as they trade off resolution for speed. Short columns, high flow rates and smaller particles are examples of these approaches. In this paper a practical method is demonstrated for injecting samples onto the column in rapid succession and gating the detection window to monitor the elution of each sample individually. At any given instant approximately two samples are eluting through the column. By co-ordinating the injection and detection time windows the samples can be kept discrete and significant throughput enhancements achieved, up to nearly 2-fold improvements are demonstrated. A rudimentary theory is development to show that the throughput improvements can be predicted to approximation by simple column characteristics. Experimental results for a series of monoclonal antibodies demonstrate the equivalency of the method to a conventional injection approach, the throughput increase, and the robustness of the method. |
| Databáze: | OpenAIRE |
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