Article (Scientific journals)
Insulin aggregation assessment by capillary gel electrophoresis without sodium dodecyl sulfate: Comparison with size-exclusion chromatography
Demelenne, Alice; NAPP, Aurore; Bouillenne, Fabrice et al.
2019In Talanta, 199, p. 457-463
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Keywords :
Acidic pH SEC; CGE; Dimers; Neutral pH SEC; Pharmacopoeia; Sodium deoxycholate; Aggregates; Amino acids; Capillary electrophoresis; Insulin; Proteins; Sodium dodecyl sulfate; Sulfur compounds; Acidic pH; Capillary gel electrophoresis; European pharmacopoeias; Mobile-phase composition; Neutral pH; Pharmacopoeia method; Size exclusion chromatography
Abstract :
[en] Size-exclusion chromatography (SEC) is a method of choice for the analysis of protein aggregates in pharmaceuticals. The United States and European Pharmacopoeias currently use a SEC method with an acidic pH mobile phase to assess the content of aggregates in insulin formulations. In this article, we analyzed aggregated human insulin samples and demonstrated that both methods under neutral conditions, namely neutral pH SEC (nSEC) and capillary gel electrophoresis (CGE), yield to similar aggregate content contrary to SEC under acidic conditions (aSEC). aSEC showed polymeric complexes that were not observed in nSEC and CGE. During method development, the effect on SEC profiles of arginine and acetonitrile were highlighted. In CGE, the effect of SDS on disruption of non-covalent insulin aggregates was confirmed and the benefit of sodium deoxycholate addition in sieving gel was discussed. The three methods were applied to the analysis of an insulin formulation and similar results to those obtained for human insulin as raw material were observed. Finally, the CGE method was used to study the stability of human insulin under different storage conditions. In view of the obtained results one may question the relevance of the current pharmacopoeia method to study insulin aggregates by emphasizing the importance of the mobile phase composition and pH in SEC. The new CGE method developed is an easy method for studying non-covalent aggregates of insulin, which could be applied to other proteins. © 2019 Elsevier B.V.
Disciplines :
Pharmacy, pharmacology & toxicology
Author, co-author :
Demelenne, Alice ;  Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments
NAPP, Aurore ;  Centre Hospitalier Universitaire de Liège - CHU > Département de pharmacie hospitalière > Service tarification, gestion des processus et stérilisation
Bouillenne, Fabrice ;  Université de Liège - ULiège > Département des sciences de la vie > Centre d'ingénierie des protéines
Crommen, Jacques ;  Université de Liège - ULiège > Département de pharmacie > Département de pharmacie
Servais, Anne-Catherine  ;  Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments
Fillet, Marianne  ;  Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments
Language :
English
Title :
Insulin aggregation assessment by capillary gel electrophoresis without sodium dodecyl sulfate: Comparison with size-exclusion chromatography
Publication date :
2019
Journal title :
Talanta
ISSN :
0039-9140
eISSN :
1873-3573
Publisher :
Elsevier B.V.
Volume :
199
Pages :
457-463
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 21 April 2019

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