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Viral clearance using monoliths.

Mark R Etzel1, William T Riordan

  • 1Department of Chemical and Biological Engineering, University of Wisconsin, 1605 Linden Drive, Madison, WI 53706, USA. Etzel@engr.wisc.edu

Journal of Chromatography. A
|October 22, 2008
PubMed
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This study evaluated salt-tolerant monoliths for removing biological impurities like viruses, DNA, and host-cell proteins (HCP) during biotechnology manufacturing. The salt-tolerant monolith demonstrated superior impurity clearance, especially at higher salt concentrations, outperforming traditional methods.

Area of Science:

  • Biotechnology Manufacturing
  • Bioprocess Engineering
  • Downstream Processing

Background:

  • Effective clearance of biological impurities is critical for biotechnology-derived products, such as monoclonal antibodies (mAbs).
  • Salt is necessary for mAb solubilization and stability but complicates impurity removal.
  • Traditional purification methods face challenges with salt interference.

Purpose of the Study:

  • To evaluate a novel salt-tolerant monolith for removing pathogenic impurities (viruses, DNA, host-cell protein [HCP]).
  • To compare its performance against a traditional quaternary amine (Q) monolith.
  • To assess the impact of salt concentration, flow rate, and impurity mixtures on clearance efficiency.

Main Methods:

  • Comparative analysis of a traditional Q monolith and a salt-tolerant monolith.

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Isolation and Genome Analysis of Single Virions using 'Single Virus Genomics'
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  • Testing impurity clearance (DNA, virus, HCP) under varying salt concentrations and flow rates.
  • Evaluation of performance with mixed impurity feed streams.

    • Main Results:

    • Both monoliths achieved DNA clearance to the limit of detection across all salt concentrations.
    • Virus and HCP clearance were comparable at zero salt.
    • The salt-tolerant monolith showed enhanced HCP clearance at intermediate salt and superior virus clearance at elevated salt concentrations.
    • Monoliths achieved high-capacity impurity trapping at significantly higher flow rates than traditional bead-based chromatography.

    Conclusions:

    • Monolith technology, particularly the salt-tolerant variant, offers an effective solution for removing diverse biological impurities.
    • The salt-tolerant monolith provides robust impurity clearance under challenging high-salt conditions common in mAb manufacturing.
    • Monoliths present a scalable and efficient alternative to traditional chromatography for biopharmaceutical purification.