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Development and scale‐up of the recovery and purification of a domain antibody Fc fusion protein‐comparison of a two and three‐step approach
Author(s) -
Herzer Sibylle,
Bhangale Atul,
Barker Gregory,
Chowdhary Isha,
Conover Matthew,
O'Mara Brian W.,
Tsang Lily,
Wang ShiYu,
Krystek Stanley R.,
Yao Yan,
Rieble Siegfried
Publication year - 2015
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.25561
Subject(s) - process development , fusion , process (computing) , leverage (statistics) , chemistry , chromatography , downstream processing , fusion protein , process engineering , computer science , biochemical engineering , recombinant dna , artificial intelligence , engineering , biochemistry , philosophy , linguistics , operating system , gene
A robust, economical process should leverage proven technology, yet be flexible enough to adopt emerging technologies which show significant benefit. Antibody and Fc‐fusion processes may capitalize on the high selectivity of an affinity capture step by reducing the total number of chromatographic steps to 2. Risk associated with this approach stems from the potentially increased time frame needed for process development as well as unforeseen changes in impurity profile during first scale‐up of drug substance (DS) for animal toxicology and clinical phase I trials (FIH) production, which could challenge a two‐step process to the point of failure. Two different purification strategies were pursued during process development for an FIH process of a dAB‐Fc fusion protein. A two‐step process was compared to a three‐step process. The two‐step process leveraged additives to maximize impurity reduction during affinity capture. While wash additives in combination with a mixed mode chromatography met all impurity reduction requirements, HCP levels were still higher as compared to the three‐step process. The three‐step process was implemented for manufacture of clinical material to mitigate risk. Biotechnol. Bioeng. 2015;112: 1417–1428. © 2015 Wiley Periodicals, Inc.