title: Effect of benzyl alcohol on recombinant human interleukin-1 receptor antagonist structure and hydrogen-deuterium exchange.
abstract: Benzyl alcohol, a preservative commonly added to multidose therapeutic protein formulations, can accelerate aggregation of recombinant human interleukin-1 receptor antagonist (rhIL-1ra). To investigate the interactions between benzyl alcohol and rhIL-1ra, we used nuclear magnetic resonance to observe the effect of benzyl alcohol on the chemical shifts of amide resonances of rhIL-1ra and to measure hydrogen-deuterium exchange rates of individual rhIL-1ra residues. Addition of 0.9% benzyl alcohol caused significant chemical shifts of amide resonances for residues 90-97, suggesting that these solvent-exposed residues participate in the binding of benzyl alcohol. In contrast, little perturbation of exchange rates was observed in the presence of either sucrose or benzyl alcohol.
authors.affiliation: Department of Chemical and Biological Engineering, University of Colorado, Boulder, Colorado 80309.
authors.forename: John R
authors.initials: JR
authors.lastname: Alford
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Fowler
authors.forename: Deborah S
authors.initials: DS
authors.lastname: Wuttke
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Kerwin
authors.forename: Ramil F
authors.initials: RF
authors.lastname: Latypov
authors.forename: John F
authors.initials: JF
authors.lastname: Carpenter
authors.forename: Theodore W
authors.initials: TW
authors.lastname: Randolph
mesh_terms: D019905:Benzyl Alcohol / Q000737:chemistry* / Q000378:metabolism; D001665:Binding Sites; D002626:Chemistry, Pharmaceutical; D041961:Deuterium Exchange Measurement; D006801:Humans; D053590:Interleukin 1 Receptor Antagonist Protein / Q000737:chemistry* / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D011310:Preservatives, Pharmaceutical / Q000737:chemistry* / Q000378:metabolism; D066329:Protein Aggregates; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry; D013329:Structure-Activity Relationship; D013678:Technology, Pharmaceutical / Q000379:methods*; D013696:Temperature
chemical_list: C507763:IL1RN protein, human; D053590:Interleukin 1 Receptor Antagonist Protein; D011310:Preservatives, Pharmaceutical; D066329:Protein Aggregates; D011994:Recombinant Proteins; D019905:Benzyl Alcohol