Effects of PEG size on structure, function and stability of PEGylated BSA

Bitten Plesner, Conan J. Fee, Peter Westh, Anders D. Nielsen

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

The effects of PEGylation on the structural, thermal and functional stability of bovine serum albumin (BSA) were investigated using BSA and 6 linear mono-PEGylated BSA compounds. The secondary and tertiary structure of BSA measured by circular dichroism (CD) was independent of PEGylation. In contrast, the thermal stability of BSA was affected by PEGylation. The apparent unfolding temperature Tmax measured by differential scanning calorimetry (DSC) decreased with PEGylation, whereas the temperature of aggregation, Tagg, measured by dynamic light scattering (DLS) increased with PEGylation. The unfolding temperature and the temperature of aggregation were both independent of the molecular weight of the PEG chain. Possible functional changes of BSA after PEGylation were measured by Isothermal Titration Calorimetry (ITC), where the binding of sodium dodecyl sulphate (SDS) to BSA and PEGylated BSA was analysed. At 25 °C, two distinct classes of binding sites (high affinity and low affinity) for BSA and one class of binding site (low affinity) for PEGylated BSA were identified. The binding isotherm was modelled assuming independence and thermodynamic equivalence of the sites within each class. From the present biophysical characterisation, it is concluded that after PEGylation BSA appears to be unaffected structurally (secondary and tertiary structure), slightly destabilised thermally (unfolding temperature), stabilised kinetically (temperature of aggregation) and has an altered functionality (binding profile). These biophysical characteristics are all independent of the molecular weight of the attached polymer chain.
OriginalsprogEngelsk
TidsskriftEuropean Journal of Pharmaceutics and Biopharmaceutics
Vol/bind79
Udgave nummer2
Sider (fra-til)399-405
ISSN0939-6411
DOI
StatusUdgivet - 2011

Emneord

  • PEGylation
  • Protein stability
  • Protein aggregation
  • Calorimetry
  • Dynamic light scattering
  • Circular dichorism

Citer dette

Plesner, Bitten ; Fee, Conan J. ; Westh, Peter ; Nielsen, Anders D. / Effects of PEG size on structure, function and stability of PEGylated BSA. I: European Journal of Pharmaceutics and Biopharmaceutics. 2011 ; Bind 79, Nr. 2. s. 399-405.
@article{317c9ce2163643cab6b3d4ec7427dc3b,
title = "Effects of PEG size on structure, function and stability of PEGylated BSA",
abstract = "The effects of PEGylation on the structural, thermal and functional stability of bovine serum albumin (BSA) were investigated using BSA and 6 linear mono-PEGylated BSA compounds. The secondary and tertiary structure of BSA measured by circular dichroism (CD) was independent of PEGylation. In contrast, the thermal stability of BSA was affected by PEGylation. The apparent unfolding temperature Tmax measured by differential scanning calorimetry (DSC) decreased with PEGylation, whereas the temperature of aggregation, Tagg, measured by dynamic light scattering (DLS) increased with PEGylation. The unfolding temperature and the temperature of aggregation were both independent of the molecular weight of the PEG chain. Possible functional changes of BSA after PEGylation were measured by Isothermal Titration Calorimetry (ITC), where the binding of sodium dodecyl sulphate (SDS) to BSA and PEGylated BSA was analysed. At 25 °C, two distinct classes of binding sites (high affinity and low affinity) for BSA and one class of binding site (low affinity) for PEGylated BSA were identified. The binding isotherm was modelled assuming independence and thermodynamic equivalence of the sites within each class. From the present biophysical characterisation, it is concluded that after PEGylation BSA appears to be unaffected structurally (secondary and tertiary structure), slightly destabilised thermally (unfolding temperature), stabilised kinetically (temperature of aggregation) and has an altered functionality (binding profile). These biophysical characteristics are all independent of the molecular weight of the attached polymer chain.",
keywords = "PEGylation, Protein stability, Protein aggregation, Calorimetry, Dynamic light scattering, Circular dichorism",
author = "Bitten Plesner and Fee, {Conan J.} and Peter Westh and Nielsen, {Anders D.}",
year = "2011",
doi = "10.1016/j.ejpb.2011.05.003",
language = "English",
volume = "79",
pages = "399--405",
journal = "European Journal of Pharmaceutics and Biopharmaceutics",
issn = "0939-6411",
publisher = "Elsevier BV",
number = "2",

}

Effects of PEG size on structure, function and stability of PEGylated BSA. / Plesner, Bitten; Fee, Conan J.; Westh, Peter; Nielsen, Anders D.

I: European Journal of Pharmaceutics and Biopharmaceutics, Bind 79, Nr. 2, 2011, s. 399-405.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Effects of PEG size on structure, function and stability of PEGylated BSA

AU - Plesner, Bitten

AU - Fee, Conan J.

AU - Westh, Peter

AU - Nielsen, Anders D.

PY - 2011

Y1 - 2011

N2 - The effects of PEGylation on the structural, thermal and functional stability of bovine serum albumin (BSA) were investigated using BSA and 6 linear mono-PEGylated BSA compounds. The secondary and tertiary structure of BSA measured by circular dichroism (CD) was independent of PEGylation. In contrast, the thermal stability of BSA was affected by PEGylation. The apparent unfolding temperature Tmax measured by differential scanning calorimetry (DSC) decreased with PEGylation, whereas the temperature of aggregation, Tagg, measured by dynamic light scattering (DLS) increased with PEGylation. The unfolding temperature and the temperature of aggregation were both independent of the molecular weight of the PEG chain. Possible functional changes of BSA after PEGylation were measured by Isothermal Titration Calorimetry (ITC), where the binding of sodium dodecyl sulphate (SDS) to BSA and PEGylated BSA was analysed. At 25 °C, two distinct classes of binding sites (high affinity and low affinity) for BSA and one class of binding site (low affinity) for PEGylated BSA were identified. The binding isotherm was modelled assuming independence and thermodynamic equivalence of the sites within each class. From the present biophysical characterisation, it is concluded that after PEGylation BSA appears to be unaffected structurally (secondary and tertiary structure), slightly destabilised thermally (unfolding temperature), stabilised kinetically (temperature of aggregation) and has an altered functionality (binding profile). These biophysical characteristics are all independent of the molecular weight of the attached polymer chain.

AB - The effects of PEGylation on the structural, thermal and functional stability of bovine serum albumin (BSA) were investigated using BSA and 6 linear mono-PEGylated BSA compounds. The secondary and tertiary structure of BSA measured by circular dichroism (CD) was independent of PEGylation. In contrast, the thermal stability of BSA was affected by PEGylation. The apparent unfolding temperature Tmax measured by differential scanning calorimetry (DSC) decreased with PEGylation, whereas the temperature of aggregation, Tagg, measured by dynamic light scattering (DLS) increased with PEGylation. The unfolding temperature and the temperature of aggregation were both independent of the molecular weight of the PEG chain. Possible functional changes of BSA after PEGylation were measured by Isothermal Titration Calorimetry (ITC), where the binding of sodium dodecyl sulphate (SDS) to BSA and PEGylated BSA was analysed. At 25 °C, two distinct classes of binding sites (high affinity and low affinity) for BSA and one class of binding site (low affinity) for PEGylated BSA were identified. The binding isotherm was modelled assuming independence and thermodynamic equivalence of the sites within each class. From the present biophysical characterisation, it is concluded that after PEGylation BSA appears to be unaffected structurally (secondary and tertiary structure), slightly destabilised thermally (unfolding temperature), stabilised kinetically (temperature of aggregation) and has an altered functionality (binding profile). These biophysical characteristics are all independent of the molecular weight of the attached polymer chain.

KW - PEGylation

KW - Protein stability

KW - Protein aggregation

KW - Calorimetry

KW - Dynamic light scattering

KW - Circular dichorism

U2 - 10.1016/j.ejpb.2011.05.003

DO - 10.1016/j.ejpb.2011.05.003

M3 - Journal article

VL - 79

SP - 399

EP - 405

JO - European Journal of Pharmaceutics and Biopharmaceutics

JF - European Journal of Pharmaceutics and Biopharmaceutics

SN - 0939-6411

IS - 2

ER -