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A label‐free methodology for selective protein quantification by means of absorption measurements
Author(s) -
Hansen Sigrid K.,
Skibsted Erik,
Staby Arne,
Hubbuch Jürgen
Publication year - 2011
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.23229
Subject(s) - throughput , bottleneck , chemistry , process (computing) , chromatography , computer science , biochemical engineering , process engineering , biological system , engineering , telecommunications , wireless , biology , embedded system , operating system
The application of high throughput experimentation (HTE) in protein purification process development has created an analytical bottleneck. Using a new label‐free and non‐invasive methodology for analyzing multicomponent protein mixtures by means of spectral measurements, we show that the analytical throughput for selective protein quantification can be increased significantly. An analytical assay based on this new methodology was shown to generate very precise results. Further, the assay was successfully applied as analytics for a resin screening performed in HTE mode. The increase in analytical throughput was obtained without decreasing the level of information when compared to analytical chromatography. This proves its potential as a valuable analytical tool in conjugation with high throughput process development (HTPD). Further, fast selective protein quantification can enhance process control in a commercial production environment and, hence, minimize the need for off‐line release analysis. Biotechnol. Bioeng. 2011;108: 2661–2669. © 2011 Wiley Periodicals, Inc.