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Vorauer-Uhl, K; Wallner, J; Lhota, G; Katinger, H; Kunert, R.
(2010): IgM characterization directly performed in crude culture supernatants by a new simple electrophoretic method
J IMMUNOL METHOD. 2010; 359(1-2): 21-27. FullText FullText_BOKU

Abstract:: A new electrophoretic technique for the qualitative and quantitative analyses of IgM isoforms and fragments has been developed. IgMs which are more complex than many other recombinantly expressed immunoglobulins are characterized by their high molecular weighted active forms and many additional isoforms and fragments in the molecular range between 25 and 1200 kDa. To analyze the multimers, isoforms and fragments simultaneously a high-resolution method, which enables sufficient migration and separation is required. Furthermore, this method should be appropriate to analyze IgMs in crude culture supernatants as well as purified samples. Simple sample preparation avoiding unspecific protein loss has been established. Currently no standard method to analyze all of them accordingly is available. The IgM-SDS-PAGE investigated for this purpose includes all these aspects. The combination of simple sample preparation and the application of precast gels make this electrophoretic method suitable for research but also quality control. The selective quantification of the multimers and the relative isoform distribution were performed by sensitive Sypro Ruby staining obtaining reliable and reproducible data in clone screening and process development which has been demonstrated by recombinantly expressed IgMs with significantly different isoform pattern. (C) 2010 Elsevier B.V. All rights reserved.
Authors BOKU Wien:: Katinger Hermann; Kunert Renate; Lhota Gabriele; Vorauer-Uhl Karola; Wallner Jakob
Find related publications in this database (using NML MeSH Indexing): Animals
CHO Cells
Cell Culture Techniques*
Cell Movement
Cell Separation
Cells, Cultured
Cricetinae
Cricetulus
Culture Media, Conditioned/chemistry*
Electrophoresis, Polyacrylamide Gel/methods*
Enzyme-Linked Immunosorbent Assay
Gels/chemistry
Immunoglobulin M/analysis*
Immunoglobulin M/immunology
Peptide Fragments/chemistry
Peptide Fragments/immunology
Protein Isoforms/chemistry
Protein Isoforms/immunology
Sensitivity and Specificity
Find related publications in this database (Keywords): IgM; Quality profiling; Quantification of IgM; Isoform distribution; IgM electrophoresis