Host cell protein quantification by fourier transform mid infrared spectroscopy (FT-MIR)

Authors

  • Florian Capito,

    Corresponding author
    1. Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Darmstadt, Germany
    2. Merck KGaA, Frankfurter Strasse 250, 64293 Darmstadt, Germany; telephone: 49-6151-727168; fax: 0049 6151 72 917510
    • Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Darmstadt, Germany
    Search for more papers by this author
  • Romas Skudas,

    1. Merck KGaA, Frankfurter Strasse 250, 64293 Darmstadt, Germany; telephone: 49-6151-727168; fax: 0049 6151 72 917510
    Search for more papers by this author
  • Harald Kolmar,

    1. Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Darmstadt, Germany
    Search for more papers by this author
  • Bernd Stanislawski

    1. Merck KGaA, Frankfurter Strasse 250, 64293 Darmstadt, Germany; telephone: 49-6151-727168; fax: 0049 6151 72 917510
    Search for more papers by this author

  • The authors declare no conflict of interest.

Abstract

Process development in up- and downstream processing requires enhanced, non-time-consuming, and non-expensive monitoring techniques to track product purity, for example, the level of endotoxins, viral particles, and host cell proteins (HCPs). Currently, HCP amounts are measured by laborious and expensive HCP-enzyme-linked immunosorbent assay (ELISA) assays best suited for measuring HCP amounts in the low concentration regime. The measurement of higher HCP amounts using this method requires dilution steps, adding dilution errors to the measurement. In this work we evaluated the suitability of attenuated total reflection spectroscopy for HCP quantification in process development, using clarified cell culture fluid from monoclonal antibody producing Chinese hamster ovary-cells after treatment with different polyelectrolytes for semi-selective clarification. Forty undiluted samples were chosen for multivariate data analysis in the middle infrared range and predicted HCP-values were in good agreement with results obtained by an ELISA-assay, suggesting the suitability of this new method for HCP-quantification. As this method is able to quantify HCP titers ranging from approximately at least 20,000–200,000 ng mL−1, it is suitable especially for monitoring of process development steps with higher HCP concentrations, omitting dilution errors associated with ELISA assays. Biotechnol. Bioeng. 2013; 110: 252–259. © 2012 Wiley Periodicals, Inc.

Ancillary