Chromatographic and electrophoretic characterization of protein variants.
Almost all proteins are expressed in several variants, also known as isoforms. Individual protein variants differ by modifications of the individual amino acid side chains, or the N- or C-terminus. Typical modifications are glycosylation, phosphorylation, acetylation, methylation, deamidation or oxidation. It is of utmost interest to either get a quantitative picture of the variants of a particular protein or to separate the variants in order to be able to identify their molecular structure. Protein variants are present in native as well as in recombinant proteins. In the case of protein production it is interesting, how variants are generated during fermentation, purification processes, storage, and how present individual variants influence the biological activity. This review provides a comparison of chromatographic and electrophoretic separation methods to analyze and to prepare protein variants.
Ahrer
Karin
K
Department of Biotechnology, University of Natural Resources and Applied Life Sciences and Austrian Center of Biopharmaceutical Technology, Vienna, Muthgasse 18, A-1190 Vienna, Austria.
Jungbauer
Alois
A
D002845:Chromatography / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods; D020374:Chromatography, Micellar Electrokinetic Capillary / Q000379:methods; D004586:Electrophoresis / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D007525:Isoelectric Focusing / Q000379:methods; D020033:Protein Isoforms / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification
D020033:Protein Isoforms