Home | english  | Impressum | KIT

High throughput screening of packed bed chromatography coupled with SELDI-TOF MS analysis: Monoclonal antibodies vs. host cell protein.

High throughput screening of packed bed chromatography coupled with SELDI-TOF MS analysis: Monoclonal antibodies vs. host cell protein.
Autor:

P.S. Wierling, R. Bogumil, E. Knieps-Grunhagen, J. Hubbuch

Links:
Quelle:

Biotechnology and Bioengineering, vol. 98, pages 440-450

Datum: 2007

A feasibility study to couple high throughput screening of packed bed chromatography with mass spectrometric detection by SELDI-TOF MS is presented. As model system monoclonal antibodies (mAb) versus host cell protein (HCP) from an industrial cultivation was chosen. Packed bed chromatography was screened on a TECAN Evo Freedom 200 station using miniaturized chromatographic columns placed on a specially designed array carrier linked to a commercially available T-Stack module. Gradient elution of the bound proteins was performed by applying a multiple step strategy. When analyzing selected HCP peaks as well as the detected antibody peaks throughout the chromatographic runs a direct correlation between applied and detected components was established. The sensitivity of conventional protein A chromatography was found to be lower than SELDI-TOF MS analysis. During initial screening a shift in the elution pattern for one of the monoclonal antibodies detected with all four resins was identified to be a heterogeneity in the mAb glycosylation pattern. In addition, a detailed differentiation between various HCP fractions through out the chromatographic process using SELDI-TOF analysis let to the detection of HCP components possibly adhering to the mAbs during chromatographic separations. Biotechnol. Bioeng. 2007;98; 440-450. © 2007 Wiley Periodicals, Inc.