Continuous precipitation of process related impurities from clarified cell culture supernatant using a novel coiled flow inversion reactor (CFIR). Issue 10 (11th August 2016)

• Record Type: Journal Article
• Authors: Kateja, Nikhil; Agarwal, Harshit; Saraswat, Aditya; Bhat, Manish; Rathore, Anurag S.
• Is Part Of: Biotechnology journal. Volume 11:Issue 10(2016)
• Page Start: 1320
• Page End: 1331
• Publication Date: 2016-08-11
• Subjects: Antibody Production -- Coiled flow inversion reactor -- Continuous precipitation -- Continuous processing -- Protein A chromatography; Biotechnology -- Periodicals; 660.605
• DOI: 10.1002/biot.201600271
• Languages: English
• ISSNs: 1860-6768

Abstract

Abstract: Coiled Flow Inverter Reactor (CFIR) has recently been explored for facilitating continuous operation of several unit operations involved in downstream processing of biopharmaceuticals such as viral inactivation and protein refolding. The application of CFIR for continuous precipitation of clarified cell culture supernatant has been explored. The pH based precipitation is optimized in the batch mode and then in the continuous mode in CFIR using a design of experiments (DOE) study. Improved clearance of host cell DNA (52× vs. 39× in batch), improved clearance of host cell proteins (HCP) (7× vs. 6× in batch) and comparable recovery (90 vs. 91.5 % in batch) are observed along with six times higher productivity. To further demonstrate wider applicability of CFIR in performing continuous precipitation, two more case studies involving use of two different precipitation protocols (CaCl2 based and caprylic acid based) are also performed. In both cases, clearance of host cell DNA, HCP, and product recovery are found to be comparable or better in CFIR than in batch operations. Moreover, increase in productivity of 16 times (CaCl2 based) and eight times (caprylic acid based) is obtained for the two precipitation protocols, respectively. The data clearly demonstrate that CFIR can be seamlessly integrated into a continuous bioprocess train for performing continuous precipitation of clarified cell culture supernatant. To our knowledge this is the first report of such use. Abstract : Various reactor configuration have been explored for continuous processing of biotech unit operations. A coiled flow inversion reactor is a novel configuration consisting of helical coils bent at equidistant right angles. This configuration provides a sharper residence time distribution in the laminar flow regime along with good cross‐sectional mixing, better emulating a plug flow than a simple straight tube or a helix. The application of this novel coiled flow inversion reactor for continuous precipitation of process related impurities from cell culture supernatant is demonstrated. This work acts as the starting point for development of an integrated continuous bioprocessing platform for therapeutics manufacturing.