Authors: Ákos Szekrényes Udo Roth Márta Kerékgyártó Andrea Székely István Kurucz Karen Kowalewski András Guttman
Publish Date: 2012/07/22
Volume: 404, Issue: 5, Pages: 1485-1494
Abstract
Capillary gel electrophoresis CGE in the presence of sodium dodecyl sulfate SDS is a wellestablished and widely used protein analysis technique in the biotechnology industry and increasingly becoming the method of choice that meets the requirements of the standards of International Conference of Harmonization ICH Automated single channel capillary electrophoresis systems are usually equipped with UV absorbance and/or laserinduced fluorescent LIF detection options offering general applicability and high detection sensitivity respectively however with limited throughput This shortcoming is addressed by the use of multicapillary gel electrophoresis mCGE systems with LEDinduced fluorescent detection LEDIF also featuring automation and excellent detection sensitivity thus widely applicable to rapid and largescale analysis of biotherapeutics especially monoclonal antibodies mAb The methodology we report in this paper is readily applicable for rapid purity assessment and subunit characterization of IgG molecules including detection of nonglycosylated heavy chains NGHC and separation of possible subunit variations such as truncated light chains PreLC or alternative splice variants Covalent fluorophore derivatization and the mCGE analysis of the labeled IgG samples with multicapillary gel electrophoresis are thoroughly described Reducing and nonreducing conditions were both applied with and without peptide Nglycosidase F mediated deglycosylation
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