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Protein PEGylation prevents benzyl‐alcohol induced aggregation
Author(s) -
RodríguezMartínez José Arcadio,
RiveraRivera Izarys,
Griebenow Kai
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.lb301
Subject(s) - chemistry , pegylation , benzyl alcohol , peg ratio , protein aggregation , steric effects , ethylene glycol , alcohol , peptide , proteolysis , organic chemistry , polyethylene glycol , biochemistry , enzyme , finance , economics , catalysis
Benzyl alcohol is currently the most commonly employed antimicrobial preservative added to protein and peptide pharmaceutical formulations. However, it has been reported that benzyl alcohol induces protein aggregation thus presenting an obstacle to safe protein formulation. Specifically, upon incubation of the model protein chymotrypsinogen at 45°C for 24 h, benzyl alcohol (0.9%) increased the formation of insoluble aggregates from 2% to 13%. We tested the effectiveness of the covalent modification of the protein with poly(ethylene glycol) (PEG) on preventing benzyl alcohol induced protein aggregation. PEGs with molecular weights (M W ) of 700 and 5000 were employed and between 2 and 5 molecules were bound to chymotrypsinogen. It was found that PEG5000 reduced aggregation to 0% while PEG 700 was largely ineffective. Since we found that there was only a marginal difference in thermodynamic stability for the various PEG‐protein conjugates, this protective effect was likely afforded by steric shielding of hydrophobic protein surface area patches. In conclusion, PEG‐modification of proteins with large PEG molecules is an effective strategy for preventing benzyl alcohol induced protein aggregation.