Highly linear pH gradients for analyzing monoclonal antibody charge heterogeneity in the alkaline range
Autor: | Muriel Bardor, Nico Lingg, Eddy Tan, Alois Jungbauer, Beate Hintersteiner |
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Přispěvatelé: | University of Natural Resources and Life Sciences (BOKU), Laboratoire de Glycobiologie et Matrice Extracellulaire Végétale (Glyco-MEV), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU), Universität für Bodenkultur Wien [Vienne, Autriche] (BOKU) |
Jazyk: | angličtina |
Rok vydání: | 2013 |
Předmět: |
[SDV.BIO]Life Sciences [q-bio]/Biotechnology
medicine.drug_class Ion chromatography Analytical chemistry [SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC] Monoclonal antibody 01 natural sciences Biochemistry Analytical Chemistry 03 medical and health sciences [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry Molecular Biology/Genomics [q-bio.GN] [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB] medicine Isoelectric Point [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biochemistry [q-bio.BM] ComputingMilieux_MISCELLANEOUS [SDV.BDD.GAM]Life Sciences [q-bio]/Development Biology/Gametogenesis 030304 developmental biology 0303 health sciences Range (particle radiation) Chromatography Chemistry 010401 analytical chemistry Organic Chemistry Linearity Antibodies Monoclonal Charge (physics) [SDV.BBM.MN]Life Sciences [q-bio]/Biochemistry Molecular Biology/Molecular Networks [q-bio.MN] [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry Molecular Biology/Molecular biology General Medicine Hydrogen-Ion Concentration Chromatography Ion Exchange 0104 chemical sciences [SDV.BV.PEP]Life Sciences [q-bio]/Vegetal Biology/Phytopathology and phytopharmacy [SDV.BV.AP]Life Sciences [q-bio]/Vegetal Biology/Plant breeding Isoelectric point [CHIM.POLY]Chemical Sciences/Polymers Immobilized pH gradient Constant (mathematics) |
Zdroj: | Journal of Chromatography A Journal of Chromatography A, Elsevier, 2013, 1319, pp.65-71. ⟨10.1016/j.chroma.2013.10.028⟩ |
ISSN: | 0021-9673 1873-3778 |
DOI: | 10.1016/j.chroma.2013.10.028⟩ |
Popis: | Recombinant antibodies with high isoelectric point are frequent since most of them are constructed from the same framework. Classically, cation exchange chromatography is used as a standard method for the determination of antibody charge heterogeneity. In contrast, in this study highly linear pH gradients were achieved by keeping the buffering capacity over the length of the gradient constant. The buffering compounds were selected to be unretained on the column and their respective concentration was adjusted in the start and end buffer of the pH gradient to achieve constant buffering capacity. This helps conserve linearity and stability of the gradient. The method allows quantification of charge variant distribution and the determination of chromatographic isoelectric point. To demonstrate the effectiveness of this novel method, a ProPac WCX-10 column was used to separate isoforms of trastuzumab biosimilar antibodies. Effects of pH gradient linearity and of varying the analytical amount of sample on the separation are shown. |
Databáze: | OpenAIRE |
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