Cation exchange chromatography provides effective retrovirus clearance for antibody purification processes

Biotechnol Bioeng. 2012 Jan;109(1):157-65. doi: 10.1002/bit.23300. Epub 2011 Aug 23.

Abstract

One measure taken to ensure safety of biotherapeutics produced in mammalian cells is to demonstrate the clearance of potential viral contaminants by downstream purification processes. This paper provides evidence that cation exchange chromatography (CEX), a widely used polishing step for monoclonal antibody (mAb) production, can effectively and reproducibly remove xMuLV, a retrovirus used as a model of non-infectious retrovirus-like particles found in Chinese hamster ovary cells. The dominant mechanism for xMuLV clearance by the strong cation exchanger, Fractogel SO ₃⁻, is by retention of the virus via adsorption instead of inactivation. Experimental data defining the design space for effective xMuLV removal by Fractogel SO ₃⁻ with respect to operational pH, elution ionic strength, loading, and load/equilibration buffer ionic strength are provided. Additionally, xMuLV is able to bind to other CEX resins, such as Fractogel COO⁻ and SP Sepharose Fast Flow, suggesting that this phenomenon is not restricted to one type of CEX resin. Taken together, the data indicate that CEX chromatography can be a robust and reproducible removal step for the model retrovirus xMuLV.

Publication types

  • Evaluation Study

MeSH terms

  • Adsorption*
  • Animals
  • Antibodies, Monoclonal / isolation & purification*
  • Biotechnology / methods*
  • Buffers
  • CHO Cells
  • Chromatography, Ion Exchange / methods*
  • Cricetinae
  • Cricetulus
  • Leukemia Virus, Murine / isolation & purification*
  • Osmolar Concentration
  • Virus Attachment*

Substances

  • Antibodies, Monoclonal
  • Buffers