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Application of calcium phosphate flocculation in high-density cell culture fluid with high product titer of monoclonal antibody

Authors
  • Chen, Guoqiang
  • Su, Zhiguo
  • Li, Feng
  • Liu, Hui Fang
Publication Date
May 01, 2017
Source
Institutional Repository of Institute of Process Engineering, CAS (IPE-IR)
Keywords
License
Unknown
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Abstract

The calcium phosphate [Ca-3(PO4)(2)] precipitation was used for improving the clarification efficiency in harvest process of the monoclonal antibody (mAb) containing cell culture fluid (CCF) with high turbidity and product titer. The flocculation conditions (concentration, addition order of flocculants, pH, and operation time), and the effect of flocculants on the mAb physical chemical properties (such as distribution of charge variants and aggregates) and process-related impurities removal (such as DNA and CHOP) were evaluated in this study. The results showed that the turbidity of CCF supernatant was significantly reduced at pH 7, 120 min with addition of phosphate ions first, while a high mAb recovery yield was kept in the CCF supernatant after flocculation. Addition of calcium ions at 15-60 mM was sufficient for flocculation in this study. A relationship between turbidity/mAb recovery yield and the concentration of calcium ions was established. More than 85% DNA in the CCF were effectively removed by the addition of optimal concentration of flocculants. Flocculation process of Ca-3(PO4)(2) is an effective pretreatment method in purification processes of mAbs from the CCF with high turbidity and product titer.

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