Enhancing Protein A performance in mAb processing: A method to reduce and rapidly evaluate host cell DNA levels during primary clarification

Biotechnol Prog. 2019 Nov;35(6):e2882. doi: 10.1002/btpr.2882. Epub 2019 Aug 13.

Abstract

The use and impact of 3M™ Emphaze™ AEX Hybrid Purifier, a single-use, fully synthetic chromatographic product, was explored to reduce host cell DNA (HC-DNA) concentration during the primary clarification of a monoclonal antibody (mAb). An approximately 5-log reduction in HC-DNA was achieved at an Emphaze AEX Hybrid Purifier throughput of 200 L/m2 . The appreciable reduction in HC-DNA achieved during primary clarification enhanced Protein A chromatography performance, resulting in a sharper and narrower elution profile. In addition, a 24× improvement in host cell protein (HCP) removal and fewer impurities nonspecifically bound to the Protein A column were observed compared to those resulting from the use of depth filtration for clarification. The use of a rapid, qualitative acidification assay to facilitate HC-DNA monitoring was also investigated. This assay involves the acidification-induced precipitation of HC-DNA, enabling the easy and rapid detection of DNA breakthrough across purification media such as Emphaze AEX Hybrid Purifier by means of turbidimetric and particle size measurements.

Keywords: DNA reduction; Emphaze AEX Hybrid Purifier; Protein A chromatography; depth filtration; turbidity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / biosynthesis*
  • CHO Cells
  • Chromatography
  • Cricetulus
  • DNA / analysis*
  • Filtration
  • Particle Size
  • Staphylococcal Protein A / chemistry*

Substances

  • Antibodies, Monoclonal
  • Staphylococcal Protein A
  • DNA