title: Separation of Recombinant Therapeutic Proteins Using Capillary Gel Electrophoresis and Capillary Isoelectric Focusing.
abstract: Detailed step-by-step methods for protein separation techniques based on capillary electrophoresis (CE) are described in this chapter. Focus is placed on two techniques, capillary gel electrophoresis (CGE) and capillary isoelectric focusing (cIEF). CGE is essentially gel electrophoresis, performed in a capillary, where a hydrogel is used as a sieving matrix to separate proteins or peptides based on size. cIEF separates proteins or peptides based on their isoelectric point (pI), the pH at which the protein or peptide bears no charges. Detailed protocols and steps (including capillary preparation, sample preparation, CE separation conditions, and detection) for both CGE and cIEF presented so that readers can follow the described methods in their own labs.
authors.affiliation: Department of Chemistry, University of British Columbia, 2036 Main Mall, Vancouver, BC, Canada, V6T 1Z1.
authors.forename: Caitlyn A G
authors.initials: CA
authors.lastname: De Jong
authors.affiliation: Department of Chemistry, University of British Columbia, 2036 Main Mall, Vancouver, BC, Canada, V6T 1Z1.
authors.forename: Jessica
authors.initials: J
authors.lastname: Risley
authors.affiliation: Department of Chemistry, University of British Columbia, 2036 Main Mall, Vancouver, BC, Canada, V6T 1Z1.
authors.forename: Alexis K
authors.initials: AK
authors.lastname: Lee
authors.affiliation: Department of Chemistry, University of British Columbia, 2036 Main Mall, Vancouver, BC, Canada, V6T 1Z1.
authors.forename: Shuai Sherry
authors.initials: SS
authors.lastname: Zhao
authors.affiliation: Department of Chemistry, University of British Columbia, 2036 Main Mall, Vancouver, BC, Canada, V6T 1Z1. chen@chem.ubc.ca.
authors.forename: David D Y
authors.initials: DD
authors.lastname: Chen
mesh_terms: D019075:Electrophoresis, Capillary / Q000379:methods*; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing / Q000379:methods*; D007526:Isoelectric Point; D011994:Recombinant Proteins / Q000302:isolation & purification*
chemical_list: D011994:Recombinant Proteins; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate
keywords: Capillary gel electrophoresis; Capillary isoelectric focusing; Protein characterization; Recombinant therapeutic proteins