title: High resolution TOF MS coupled to CE for the analysis of isotopically resolved intact proteins.
abstract: Intact protein analysis by mass spectrometry is of great interest for the characterisation of biotechnological products. Exact mass measurement in combination with isotopic resolution allows the detection of modifications leading to small mass changes like deamidation or reduction of disulfide bonds directly on the level of the intact protein. Here, a concept is presented based on time-of-flight mass spectrometry. A bench top TOF MS and a high resolution TOF MS were used to resolve the isotopes of intact recombinant human growth hormone and intact human erythropoietin, respectively. Thus, these 22 and around 30kDa large proteins can be characterised sensitively in great detail and along with capillary electrophoretic separation unambiguous identification of minor protein modifications like deamidation is possible.
authors.affiliation: Aalen University, Aalen, Germany.
authors.forename: Angelina
authors.initials: A
authors.lastname: Taichrib
authors.forename: Matthias
authors.initials: M
authors.lastname: Pelzing
authors.forename: Cristoforo
authors.initials: C
authors.lastname: Pellegrino
authors.forename: Mara
authors.initials: M
authors.lastname: Rossi
authors.forename: Christian
authors.initials: C
authors.lastname: Neusüss
mesh_terms: D019075:Electrophoresis, Capillary / Q000379:methods; D004921:Erythropoietin / Q000032:analysis; D019382:Human Growth Hormone / Q000032:analysis*; D006801:Humans; D008970:Molecular Weight; D020033:Protein Isoforms / Q000302:isolation & purification; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000032:analysis* / Q000378:metabolism; D011994:Recombinant Proteins / Q000032:analysis; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
chemical_list: D020033:Protein Isoforms; D011506:Proteins; D011994:Recombinant Proteins; D004921:Erythropoietin; D019382:Human Growth Hormone