Self-association of long-acting insulin analogues studied by size exclusion chromatography coupled to multi-angle light scattering

Two structurally very different insulin analogues analysed here, belong to a class of analogues of which two have been reported to have a protracted action through self-assembly to high molar mass in subcutis. The process of self-association of insulin analogues Lys B29 (N ɛω-carboxyheptadecanoyl) d...

Full description

Saved in:
Bibliographic Details
Published in:Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2011-10, Vol.879 (28), p.2945-2951
Main Authors: Jensen, Malene H., Wahlund, Per-Olof, Jacobsen, Jes K., Vestergaard, Bente, van de Weert, Marco, Havelund, Svend
Format: Article
Language:English
Subjects:
Acylated insulin analogue
Amino Acid Sequence
Analysis
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Chromatography, Gel - methods
Depot formation
Formulations
Fundamental and applied biological sciences. Psychology
gel chromatography
General pharmacology
Human
Humans
Insulin
Insulin, Long-Acting - chemistry
Kinetics
Light scattering
Long-acting insulin analogue
Medical sciences
Molecular Sequence Data
Molecular Structure
molecular weight
Pharmacology. Drug treatments
phenol
preservatives
Scattering, Radiation
Self assembly
Self-association of insulin analogues
Sheds
Size exclusion chromatography
Size exclusion chromatography coupled with multi-angle light-scattering
Soluble basal insulin
Zinc
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Two structurally very different insulin analogues analysed here, belong to a class of analogues of which two have been reported to have a protracted action through self-assembly to high molar mass in subcutis. The process of self-association of insulin analogues Lys B29 (N ɛω-carboxyheptadecanoyl) des(B30) human insulin and Lys B29 (N ɛ-lithocholyl) des(B30) human insulin was investigated using size exclusion chromatography (SEC) in connection with multi-angle light-scattering. Self-assembly to high molar mass was obtained by exchanging the formulation containing phenolic preservatives with an isotonic eluent during SEC. It was shown that increasing amounts of zinc in the formulations of the two analogues increased the size of the self assemblies formed during gel filtration. The addition of 0.2 mM phenol to the elution buffer slowed down the self-association process of zinc containing formulations and shed light on the initial association process. The results indicated that a dihexamer is a possible building block during self-association of Lys B29 (N ɛω-carboxyheptadecanoyl) des(B30) human insulin. Surprisingly, in the absence of zinc the two analogues behaved very differently. Lys B29 (N ɛω-carboxyheptadecanoyl) des(B30) human insulin was in equilibrium between oligomers smaller than a hexamer, whereas Lys B29 (N ɛ-lithocholyl) des(B30) human insulin self-associated and formed even larger complexes than in the presence of zinc.
ISSN:1570-0232
1873-376X
DOI:10.1016/j.jchromb.2011.05.059