Mass Spectrometric Analysis of Intact Human Monoclonal Antibody Aggregates Fractionated by Size-Exclusion Chromatography

• DOI: 10.1007/s11095-010-0224-5
• Publication Date: 2010-08-02T19:58:34Z
• Authors: Başak Kükrer; Vasco Filipe; Esther van Duijn; Piotr T. Kasper; Rob J. Vreeken; Albert J. R. Heck; Wim Jiskoot
• Keywords: Oligomer; Molecular mass

The aim of this study was to develop a method characterize intact soluble monoclonal IgG1 antibody (IgG) oligomers by mass spectrometry.IgG aggregates (dimers, trimers, tetramers and high-molecular-weight oligomers) were created subjecting an IgG formulation several pH jumps. Protein oligomer fractions isolated high performance size exclusion chromatography (HP-SEC), dialyzed against ammonium acetate 6.0 (a spectrometry-compatible volatile buffer), analyzed native electrospray ionization time-of-flight spectrometry (ESI-TOF MS).Monomeric aggregated in the stressed successfully HP-SEC. ESI-TOF MS analysis enabled us determine molecular weight monomeric as well aggregates, including dimers, trimers tetramers. HP-SEC separation sample preparation proved be necessary for good quality signal MS. Both protocol spectrometric technique shown leave largely intact.ESI-TOF is useful tool complementary identify small oligomeric protein aggregates.