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      Continuous precipitation of process related impurities from clarified cell culture supernatant using a novel coiled flow inversion reactor (CFIR).

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          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.

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          Author and article information

          Journal
          Biotechnol J
          Biotechnology journal
          Wiley
          1860-7314
          1860-6768
          Oct 2016
          : 11
          : 10
          Affiliations
          [1 ] Department of Chemical Engineering, Indian Institute of Technology Delhi, New Delhi, India.
          [2 ] Department of Chemical Engineering, Indian Institute of Technology Delhi, New Delhi, India. asrathore@biotechcmz.com.
          Article
          10.1002/biot.201600271
          27447837
          586e160c-3d78-4932-a9f4-fb0a84e1b2c6
          History

          Coiled flow inversion reactor,Continuous processing,Protein A chromatography,Continuous precipitation,Antibody Production

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