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A tandem laboratory scale protein purification process using Protein A affinity and anion exchange chromatography operated in a weak partitioning mode
Author(s) -
Shamashkin Michael,
Godavarti Ranga,
Iskra Timothy,
Coffman Jon
Publication year - 2013
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.24955
Subject(s) - tandem , fast protein liquid chromatography , process engineering , cross flow filtration , chromatography , diafiltration , chemistry , downstream processing , tandem affinity purification , unit operation , process (computing) , batch processing , computer science , affinity chromatography , materials science , chemical engineering , high performance liquid chromatography , microfiltration , engineering , membrane , programming language , composite material , enzyme , operating system , biochemistry
A significant consequence of scaling up production of high titer monoclonal antibody (mAb) processes in existing facilities is the generation of in‐process pools that exceed the capacity of storage vessels. A semi‐continuous downstream process where columns and filters are linked and operated in tandem would eliminate the need for intermediate holding tanks. This study is a bench‐scale demonstration of the feasibility of a tandem process for the purification of mAbs employing an affinity Protein A capture step, followed by a flow‐through anion‐exchange (AEX) step with the possibility of adding an in‐line virus filtration step (VF). All three steps were linked sequentially and operated as one continuous process using an ÄKTA FPLC equipped with two pumps and a system of valves and bypasses that allowed the components to be engaged at different stages of the process. The AEX column was operated in a weak partitioning (WP) mode enabled by a precise in‐line titration of Protein A effluent. In order to avoid complex control schemes and facilitate validation, quality and robustness were built into the system through selection of buffers based on thermodynamic and empirical models. The tandem system utilized the simplest possible combination of valves, pumps, controls, and automation, so that it could easily be implemented in a clinical or commercial production facility. Linking the purification steps in a tandem process is expected to generate savings in time and production costs and also reduce the size of quality systems due to reduced documentation requirements, microbial sampling, and elimination of hold time validation. Biotechnol. Bioeng. 2013;110: 2655–2663. © 2013 Wiley Periodicals, Inc.