Influence of Acylation on the Adsorption of Insulin to Hydrophobic Surfaces

Pinholt, Charlotte; Hostrup, Susanne; Bukrinsky, Jens T.; Frøkjær, Sven; Jørgensen, Lene

doi:10.1007/s11095-010-0349-6

Cited by 15 publications

(15 citation statements)

References 37 publications

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“…For acylated insulin, the association behavior (ability to form hexamer structures) and the adsorption to hydrophobic surfaces was modified. When insulin was mainly monomers and dimers, more of the acylated insulin adsorbed to the surface, but when insulin and acylated insulin was a mixture of monomer, dimer and hexamer, no differences were observed [164]. Altering the hydrophobicity of a protein can also change its surface activity; the surface activity of IgG increased when it was modified by covalently binding a C8-C16 fatty acid [87], which will also influence its adsorption behavior.…”

Section: Effects Of Modification Of Proteinsmentioning

confidence: 99%

The importance of interfaces in protein drug delivery – why is protein adsorption of interest in pharmaceutical formulations?

Pinholt¹,

Hartvig²,

Medlicott

et al. 2011

Expert Opinion on Drug Delivery

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Section: Effects Of Modification Of Proteinsmentioning

confidence: 99%

The importance of interfaces in protein drug delivery – why is protein adsorption of interest in pharmaceutical formulations?

Pinholt¹,

Hartvig²,

Medlicott

et al. 2011

Expert Opinion on Drug Delivery

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110

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“…As a pharmaceutical tool, lipidation of peptides and proteins can be used to increase the circulation time in the bloodstream, and thus increase the therapeutic efficacy of biomolecular compounds [4][5][6] . However, modifying soluble proteins with a hydrophobic moiety such as a lipid tail will inevitably have an effect on the physiochemical properties of the protein, including its physical stability [7][8][9][10][11] . Moreover, protein drugs encounter several surfaces and interfaces during the entire lifetime of the compounds: from production, purification and storage to administration, delivery and in vivo utilization.…”

Section: Introductionmentioning

confidence: 99%

Lipidation Effect on Surface Adsorption and Associated Fibrillation of the Model Protein Insulin

et al. 2016

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Citation for published version (APA):Fogh Hedegaard, S., Cardenas, M., Barker, R., Jorgensen, L., & van der Weert, M. (2016). Lipidation effect on surface adsorption and associated fibrillation of the model protein insulin. Langmuir, 32(28), 7241-7249. DOI: 10.1021/acs.langmuir.6b00522 General rightsCopyright and moral rights for the publications made accessible in Discovery Research Portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights.• Users may download and print one copy of any publication from Discovery Research Portal for the purpose of private study or research.• You may not further distribute the material or use it for any profit-making activity or commercial gain.• You may freely distribute the URL identifying the publication in the public portal. Take down policyIf you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. ABSTRACTLipidation of proteins is used in the pharmaceutical field to increase the therapeutic efficacy of proteins. In this study, we investigate the effect of a 14-carbon fatty acid modification on the adsorption behavior of human insulin to a hydrophobic solid surface and the subsequent fibrillation development under highly acidic conditions and elevated temperature by comparing to the fibrillation of human insulin. At these stressed conditions, the lipid modification accelerates the rate of fibrillation in bulk solution. With the use of several complementary surface-sensitive techniques, including quartz crystal microbalance with dissipation monitoring (QCM-D), atomic force microscopy (AFM) and neutron reflectivity (NR), we show that there are two levels of structurally different protein organization at a hydrophobic surface for both human insulin and the lipidated analogue: a dense protein layer formed within minutes on the surface and a diffuse outer layer of fibrillar structures which took hours to form. The two layers may only be weakly connected and proteins from both layers are able to desorb from the surface. The lipid modification increases the protein surface coverage and the thickness of both layer organizations. 2Upon lipidation not only the fibrillation extent but also the morphology of the fibrillar structures changes from fibril clusters on the surface to a more homogenous network of fibrils covering the entire hydrophobic surface.

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“…Most detailed structural studies have been carried out at physiological pH, from which the following picture of insulin adsorption emerges. Insulin monomers often form a close-packed monolayer at hydrophobic interfaces, where they lose α-helical structure and gain random coil or β-sheet structure [10][11][12][13][14] . Conformational changes in adsorbed insulin appear to allow further binding from solution, such that pre-adsorbed peptides with a β-sheet structure were found to accelerate surface-induced insulin fibrillation 15,16 .…”

Section: Introductionmentioning

confidence: 99%

“…The expectation from the literature for the secondary structure of adsorbed insulin is a decrease in α-helical content and an increase in β-sheet or random coil. This was deduced from UV circular dichroism spectra on Teflon and polystyrene beads at pH 7.410,13,14 , while an FTIR-ATR study of insulin adsorbed on a phenyldimethyl-functionalised silicon crystal interpreted a peak at 1708 cm…”

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Stabilization of Insulin by Adsorption on a Hydrophobic Silane Self-Assembled Monolayer

et al. 2015

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The interaction between many proteins and hydrophobic functionalized surfaces is known to induce β-sheet and amyloid fibril formation. In particular, insulin has served as a model peptide to understand such fibrillation, but the early stages of insulin misfolding and the influence of the surface have not been followed in detail under the acidic conditions relevant to the synthesis and purification of insulin. Here we compare the adsorption of human insulin on a hydrophobic (-CH3-terminated) silane self-assembled monolayer to a hydrophilic (-NH3(+)-terminated) layer. We monitor the secondary structure of insulin with Fourier transform infrared attenuated total reflection and side-chain orientation with sum frequency spectroscopy. Adsorbed insulin retains a close-to-native secondary structure on both hydrophobic and hydrophilic surfaces for extended periods at room temperature and converts to a β-sheet-rich structure only at elevated temperature. We propose that the known acid stabilization of human insulin and the protection of the aggregation-prone hydrophobic domains on the insulin monomer by adsorption on the hydrophobic surface work together to inhibit fibril formation at room temperature.

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Influence of Acylation on the Adsorption of Insulin to Hydrophobic Surfaces

Abstract: The influence of acylation on the adsorption behavior of insulin depends on the association degree of insulin, possibly due to a greater difference in hydrophobicity between monomeric insulin and acylated insulin than between the hexameric forms of these two proteins.

Cited by 15 publications

References 37 publications

The importance of interfaces in protein drug delivery – why is protein adsorption of interest in pharmaceutical formulations?

The importance of interfaces in protein drug delivery – why is protein adsorption of interest in pharmaceutical formulations?

Lipidation Effect on Surface Adsorption and Associated Fibrillation of the Model Protein Insulin

Stabilization of Insulin by Adsorption on a Hydrophobic Silane Self-Assembled Monolayer

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