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High‐throughput screening of packed‐bed chromatography coupled with SELDI‐TOF MS analysis: monoclonal antibodies versus host cell protein
Author(s) -
Wierling Pierre Schulze,
Bogumil Ralf,
KniepsGrünhagen Esther,
Hubbuch Jürgen
Publication year - 2007
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.21399
Subject(s) - chromatography , monoclonal antibody , chemistry , elution , mass spectrometry , antibody , biology , immunology
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.