title: 2-DE and MALDI-TOF-MS analysis of therapeutic fusion protein abatacept.
abstract: 2-DE and MALDI-TOF MS are useful techniques for the quality evaluation of medicinal products derived from recombinant DNA technology. The principal objective of this study has been to evaluate the suitability of 2-DE in combination with MALDI-TOF MS for the quality study of the therapeutic recombinant protein, abatacept. 1-DE SDS-PAGE, under reducing and nonreducing conditions, and 2-DE analysis were used for the assessment of M(r) , pI, and enzymatic deglycosylation efficiency of abatacept. 2-DE allowed the assessment of product identity, purity, charge heterogeneity, isoform pattern, and post-translational modifications. Furthermore, optimization of the deglycosylation procedure, charge heterogeneity, and sample preparation for the subsequent MALDI-TOF MS analysis has been addressed. PMF analysis allowed rapid identity confirmation of abatacept.
authors.affiliation: Department of Medicinal Chemistry, Faculty of Life Sciences, University of Vienna, Vienna, Austria.
authors.forename: Dashnor
authors.initials: D
authors.lastname: Nebija
authors.forename: Ernst
authors.initials: E
authors.lastname: Urban
authors.forename: Martina
authors.initials: M
authors.lastname: Stessl
authors.forename: Christian R
authors.initials: CR
authors.lastname: Noe
authors.forename: Bodo
authors.initials: B
authors.lastname: Lachmann
mesh_terms: D000069594:Abatacept; D000595:Amino Acid Sequence; D016208:Databases, Factual; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006031:Glycosylation; D018796:Immunoconjugates / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D018796:Immunoconjugates; D011993:Recombinant Fusion Proteins; D000069594:Abatacept