title: Quality control and stability studies with the monoclonal antibody, trastuzumab: application of 1D- vs. 2D-gel electrophoresis.
abstract: Recombinant monoclonal antibodies (rmAbs) are medicinal products obtained by rDNA technology. Consequently, like other biopharmaceuticals, they require the extensive and rigorous characterization of the quality attributes, such as identity, structural integrity, purity and stability. The aim of this work was to study the suitability of gel electrophoresis for the assessment of charge heterogeneity, post-translational modifications and the stability of the therapeutic, recombinant monoclonal antibody, trastuzumab. One-dimensional, SDS-PAGE, under reducing and non-reducing conditions, and two-dimensional gel electrophoresis were used for the determination of molecular mass (Mr), the isoelectric point (pI), charge-related isoform patterns and the stability of trastuzumab, subjected to stressed degradation and long-term conditions. For the assessment of the influence of glycosylation in the charge heterogeneity pattern of trastuzumab, an enzymatic deglycosylation study has been performed using N-glycosidase F and sialidase, whereas carboxypeptidase B was used for the lysine truncation study. Experimental data documented that 1D and 2D gel electrophoresis represent fast and easy methods to evaluate the quality of biological medicinal products. Important stability parameters, such as the protein aggregation, can be assessed, as well.
authors.affiliation: Department of Pharmaceutical Chemistry, Medical Faculty, Rr. Bulevardi i Deshmoreve, n.n. 10000 Pristina, Kosovo. dashnor.nebija@uni-pr.edu.
authors.forename: Dashnor
authors.initials: D
authors.lastname: Nebija
authors.affiliation: Department of Pharmaceutical Chemistry, Faculty of Life Sciences, University of Vienna, Althanstrasse 14, 1090 Vienna, Austria. christian.noe@univie.ac.at.
authors.forename: Christian R
authors.initials: CR
authors.lastname: Noe
authors.affiliation: Department of Pharmaceutical Chemistry, Faculty of Life Sciences, University of Vienna, Althanstrasse 14, 1090 Vienna, Austria. ernst.urban@univie.ac.at.
authors.forename: Ernst
authors.initials: E
authors.lastname: Urban
authors.affiliation: Department of Pharmaceutical Chemistry, Faculty of Life Sciences, University of Vienna, Althanstrasse 14, 1090 Vienna, Austria. bodo.lachmann@univie.ac.at.
authors.forename: Bodo
authors.initials: B
authors.lastname: Lachmann
mesh_terms: D000911:Antibodies, Monoclonal / Q000032:analysis*; D061067:Antibodies, Monoclonal, Humanized / Q000032:analysis*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000592:standards; D004591:Electrophoresis, Polyacrylamide Gel / Q000592:standards; D006031:Glycosylation; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008970:Molecular Weight; D020033:Protein Isoforms / Q000032:analysis; D055550:Protein Stability; D011786:Quality Control; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D013696:Temperature; D000068878:Trastuzumab
chemical_list: D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D020033:Protein Isoforms; D011994:Recombinant Proteins; D000068878:Trastuzumab
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