title: Capillary zone electrophoresis method for a highly glycosylated and sialylated recombinant protein: development, characterization and application for process development.
abstract: A purity method based on capillary zone electrophoresis (CZE) has been developed for the separation of isoforms of a highly glycosylated protein. The separation was found to be driven by the number of sialic acids attached to each isoform. The method has been characterized using orthogonal assays and shown to have excellent specificity, precision and accuracy. We have demonstrated the CZE method is a useful in-process assay to support cell culture and purification development of this glycoprotein. Compared to isoelectric focusing (IEF), the CZE method provides more quantitative results and higher sample throughput with excellent accuracy, qualities that are required for process development. In addition, the CZE method has been applied in the stability testing of purified glycoprotein samples.
authors.affiliation: Analytical Sciences of Drug Substance Department, Amgen, Inc. , Thousand Oaks, California 91320, United States.
authors.forename: Le
authors.initials: L
authors.lastname: Zhang
authors.forename: Ken
authors.initials: K
authors.lastname: Lawson
authors.forename: Bernice
authors.initials: B
authors.lastname: Yeung
authors.forename: Jette
authors.initials: J
authors.lastname: Wypych
mesh_terms: D001681:Biological Assay / Q000379:methods*; D002478:Cells, Cultured; D019075:Electrophoresis, Capillary / Q000379:methods*; D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods; D019158:N-Acetylneuraminic Acid / Q000032:analysis* / Q000737:chemistry; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry; D020033:Protein Isoforms; D011994:Recombinant Proteins / Q000032:analysis* / Q000737:chemistry
chemical_list: D006023:Glycoproteins; D011134:Polysaccharides; D020033:Protein Isoforms; D011994:Recombinant Proteins; D019158:N-Acetylneuraminic Acid