title: Characterization of a biopharmaceutical protein and evaluation of its purification process using automated capillary Western blot.
abstract: This paper describes the application of an automated size-based capillary Western blot system (Sally instrument) from ProteinSimple, Inc., for biopharmaceutical fusion-Fc protein characterization and evaluation of its purification process. The fusion-Fc protein column purification from an excess of single chain Fc polypeptide and removal of an enzyme coexpressed for protein maturation have been demonstrated using an automated capillary Western system. The clearance of a selected host cell protein (HCP) present in cell culture of fusion-Fc protein was also quantitatively monitored throughout the protein purification process. Additionally, the low levels of fusion-Fc product-related impurities detected by traditional slab gel Western blot were confirmed by the automated capillary Western system. Compared to the manual approach, the automated capillary Western blot provides the advantages of ease of operation, higher sample throughput, greater linearity range, and higher precision for protein quantitation.
authors.affiliation: Biogen Idec Inc, Cambridge, MA, USA.
authors.forename: Dong
authors.initials: D
authors.lastname: Xu
authors.forename: Sarthak
authors.initials: S
authors.lastname: Mane
authors.forename: Zoran
authors.initials: Z
authors.lastname: Sosic
mesh_terms: D001331:Automation; D015153:Blotting, Western / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods*; D004798:Enzymes / Q000302:isolation & purification; D018840:HSP70 Heat-Shock Proteins / Q000302:isolation & purification; D007141:Immunoglobulin Fc Fragments / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification*
chemical_list: D004798:Enzymes; D018840:HSP70 Heat-Shock Proteins; D007141:Immunoglobulin Fc Fragments; D011993:Recombinant Fusion Proteins
keywords: Automated Western blot; Biopharmaceutical protein characterization; CE; HCP clearance; Purification process evaluation