The authors declare no conflict of interest.
Host cell protein quantification by fourier transform mid infrared spectroscopy (FT-MIR)†
Article first published online: 8 AUG 2012
Copyright © 2012 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 110, Issue 1, pages 252–259, January 2013
How to Cite
Capito, F., Skudas, R., Kolmar, H. and Stanislawski, B. (2013), Host cell protein quantification by fourier transform mid infrared spectroscopy (FT-MIR). Biotechnol. Bioeng., 110: 252–259. doi: 10.1002/bit.24611
- Issue published online: 20 NOV 2012
- Article first published online: 8 AUG 2012
- Accepted manuscript online: 18 JUL 2012 10:51AM EST
- Manuscript Accepted: 11 JUL 2012
- Manuscript Received: 11 JUN 2012
- Merck KGaA
- host cell proteins;
- attenuated total reflection;
- Fourier transform infrared spectroscopy;
- partial least squares regression;
- process development;
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.