The Effect of Protein PEGylation on Physical Stability in Liquid Formulation
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The Effect of Protein PEGylation on Physical Stability in Liquid Formulation. / Holm, Louise Stenstrup; Mcumber, Aaron; Rasmussen, Jakob Ewald; Obiols-Rabasa, Marc; Thulstrup, Peter W.; Kasimova, Marina R.; Randolph, Theodore W.; van de Weert, Marco.
In: Journal of Pharmaceutical Sciences, Vol. 103, No. 10, 21.08.2014, p. 3043-3054.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - The Effect of Protein PEGylation on Physical Stability in Liquid Formulation
AU - Holm, Louise Stenstrup
AU - Mcumber, Aaron
AU - Rasmussen, Jakob Ewald
AU - Obiols-Rabasa, Marc
AU - Thulstrup, Peter W.
AU - Kasimova, Marina R.
AU - Randolph, Theodore W.
AU - van de Weert, Marco
PY - 2014/8/21
Y1 - 2014/8/21
N2 - The presence of micron aggregates in protein formulations has recently attracted increased interest from regulatory authorities, industry, and academia because of the potential undesired side effects of their presence. In this study, we characterized the micron aggregate formation of hen egg-white lysozyme (Lyz) and its diPEGylated (5 kDa) analog as a result of typical handling stress conditions. Both proteins were subjected to mechanical stress in the absence and presence of silicone oil (SO), elevated temperatures, and freeze-thaw cycles. Flow imaging microscopy showed that PEGylated Lyz formed approximately half as many particles as Lyz, despite its lower apparent thermodynamic stability and more loose protein fold. Further characterization showed that the PEGylation led to a change from attractive to repulsive protein-protein interactions, which may partly explain the reduced particle formation. Surprisingly, the PEGylated Lyz adsorbed an order of magnitude faster onto SO, despite being much larger in size, as determined by small-angle X-ray scattering and dynamic light scattering measurements. Thus, PEGylation may significantly reduce, but not prevent, micron aggregate formation of a protein during typical handling stresses.
AB - The presence of micron aggregates in protein formulations has recently attracted increased interest from regulatory authorities, industry, and academia because of the potential undesired side effects of their presence. In this study, we characterized the micron aggregate formation of hen egg-white lysozyme (Lyz) and its diPEGylated (5 kDa) analog as a result of typical handling stress conditions. Both proteins were subjected to mechanical stress in the absence and presence of silicone oil (SO), elevated temperatures, and freeze-thaw cycles. Flow imaging microscopy showed that PEGylated Lyz formed approximately half as many particles as Lyz, despite its lower apparent thermodynamic stability and more loose protein fold. Further characterization showed that the PEGylation led to a change from attractive to repulsive protein-protein interactions, which may partly explain the reduced particle formation. Surprisingly, the PEGylated Lyz adsorbed an order of magnitude faster onto SO, despite being much larger in size, as determined by small-angle X-ray scattering and dynamic light scattering measurements. Thus, PEGylation may significantly reduce, but not prevent, micron aggregate formation of a protein during typical handling stresses.
KW - Calorimetry (DSC)
KW - Circular dichroism
KW - Light scattering
KW - Microflow imaging
KW - PEGylation
KW - Physical stability
KW - Protein aggregation
KW - Proteins
KW - Silicone oil
KW - Small-angle X-ray scattering
U2 - 10.1002/jps.24094
DO - 10.1002/jps.24094
M3 - Journal article
C2 - 25139193
VL - 103
SP - 3043
EP - 3054
JO - Journal of Pharmaceutical Sciences
JF - Journal of Pharmaceutical Sciences
SN - 0022-3549
IS - 10
ER -
ID: 123777410