Premium
Characterization of a biopharmaceutical protein and evaluation of its purification process using automated capillary Western blot
Author(s) -
Xu Dong,
Mane Sarthak,
Sosic Zoran
Publication year - 2015
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201400380
Subject(s) - biopharmaceutical , characterization (materials science) , chromatography , western blot , capillary electrophoresis , process (computing) , capillary action , chemistry , materials science , nanotechnology , biochemistry , computer science , biology , microbiology and biotechnology , gene , composite material , operating system
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.