title: Continuous precipitation of process related impurities from clarified cell culture supernatant using a novel coiled flow inversion reactor (CFIR).
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 (CaCl2based 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 (CaCl2based) 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.
authors.affiliation: Department of Chemical Engineering, Indian Institute of Technology Delhi, New Delhi, India.
authors.forename: Nikhil
authors.initials: N
authors.lastname: Kateja
authors.affiliation: Department of Chemical Engineering, Indian Institute of Technology Delhi, New Delhi, India.
authors.forename: Harshit
authors.initials: H
authors.lastname: Agarwal
authors.affiliation: Department of Chemical Engineering, Indian Institute of Technology Delhi, New Delhi, India.
authors.forename: Aditya
authors.initials: A
authors.lastname: Saraswat
authors.affiliation: Department of Chemical Engineering, Indian Institute of Technology Delhi, New Delhi, India.
authors.forename: Manish
authors.initials: M
authors.lastname: Bhat
authors.affiliation: Department of Chemical Engineering, Indian Institute of Technology Delhi, New Delhi, India. asrathore@biotechcmz.com.
authors.forename: Anurag S
authors.initials: AS
authors.lastname: Rathore
mesh_terms: D000818:Animals; D019149:Bioreactors; D001709:Biotechnology / Q000295:instrumentation* / Q000379:methods; D016466:CHO Cells; D002122:Calcium Chloride / Q000737:chemistry; D002210:Caprylates / Q000737:chemistry; D018929:Cell Culture Techniques; D011232:Chemical Precipitation; D003412:Cricetulus; D004247:DNA / Q000032:analysis*; D011506:Proteins / Q000032:analysis*
chemical_list: D002210:Caprylates; D011506:Proteins; D004247:DNA; D002122:Calcium Chloride; C031492:octanoic acid
keywords: Antibody Production; Coiled flow inversion reactor; Continuous precipitation; Continuous processing; Protein A chromatography