Negative gradient slope methods to improve the separation of closely eluting proteins.
gradient elution
method development
monoclonal antibody
negative gradient step
negative segmented multi-isocratic mode
therapeutic proteins
Journal
Journal of chromatography. A
ISSN: 1873-3778
Titre abrégé: J Chromatogr A
Pays: Netherlands
ID NLM: 9318488
Informations de publication
Date de publication:
04 Jan 2021
04 Jan 2021
Historique:
received:
26
08
2020
revised:
16
11
2020
accepted:
19
11
2020
pubmed:
2
12
2020
medline:
1
1
2021
entrez:
1
12
2020
Statut:
ppublish
Résumé
In the present work, we describe the fundamental and practical advantages of a new strategy to improve the resolution of very closely eluting peaks within therapeutic protein samples. This approach involves the use of multiple isocratic steps, together with the addition of a steep negative gradient segment (with a decrease in mobile phase strength) to "park" a slightly more retained peak somewhere along the column (at a given migration distance), while a slightly less retained compound can be eluted. First, some model calculations were performed to highlight the potential of this innovative approach. For this purpose, the retention parameters (logk
Identifiants
pubmed: 33260022
pii: S0021-9673(20)31017-7
doi: 10.1016/j.chroma.2020.461743
pii:
doi:
Substances chimiques
Antibodies, Monoclonal
0
Indicators and Reagents
0
Proteins
0
Solutions
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
461743Informations de copyright
Copyright © 2020. Published by Elsevier B.V.
Déclaration de conflit d'intérêts
Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.