title: Glycation of interferon-beta-1b and human serum albumin in a lyophilized glucose formulation. Part III: application of proteomic analysis to the manufacture of biological drugs.
abstract: Glycation of interferon-beta-1b and human serum albumin was identified in a lyophilized glucose formulation by a sensitive LC-MS approach. The extent of glycation was measured by a label-free quantitation strategy. The glycation sites were determined by the accurate mass (FTICR MS) with MS/MS measurements on the corresponding tryptic peptides. The extent of glycation was measured by the ratio of the peak intensity between the glycated and the average value for three non-glycated peptides in the same run. Residues lysine 18 of interferon-beta-1b, and lysine 51, lysine 233, and lysine 545 of human serum albumin were more prone to be glycated than other sites in this lyophilized glucose formulation. Residues of lysine 51 and lysine 233 but not lysine 545 of human serum albumin are highly accessible to solvent as found in a solution storage study by Lapolla et al. The extent of glycation of both proteins and the number of glycation sites of human serum albumin were increased with the storage time at 25 degrees C. In total, two glycation sites of interferon beta-1b and 17 glycation sites of human serum albumin were identified in the lyophilized glucose formulation with a storage time at 25 degrees C of 35 days. Among the 17 glycation sites, only lysine 525 of human serum albumin has been found in vivo in diabetic patients by Shaklai et al. As expected, there was no glycation found on both interferon-beta-1b and human serum albumin in the control samples (similar lyophilized formulation but using mannitol instead of glucose).
authors.affiliation: Barnett Institute, Department of Chemistry and Chemical Biology, Northeastern University, 341 Mugar Hall, 360 Huntington Avenue, Boston, MA 02115, United States.
authors.forename: Xiaoyang
authors.initials: X
authors.lastname: Zheng
authors.forename: Shiaw-Lin
authors.initials: SL
authors.lastname: Wu
authors.forename: William S
authors.initials: WS
authors.lastname: Hancock
mesh_terms: D000595:Amino Acid Sequence; D002853:Chromatography, Liquid; D005947:Glucose / Q000737:chemistry*; D006031:Glycosylation; D006801:Humans; D000068576:Interferon beta-1b; D016899:Interferon-beta / Q000737:chemistry*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D040901:Proteomics; D011994:Recombinant Proteins / Q000737:chemistry; D012709:Serum Albumin / Q000737:chemistry*
chemical_list: D010455:Peptides; D011994:Recombinant Proteins; D012709:Serum Albumin; D000068576:Interferon beta-1b; D016899:Interferon-beta; D005947:Glucose