Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 428008
Title Multi-dimensional fractionation and characterization of crude protein mixtures: Toward establishment of a database of protein purification process development parameters
Author(s) Ahamed, T.; Pinkse, M.W.H.; Wielen, L.A.M. van der; Verhaert, P.; Dedem, G.; Eppink, M.H.M.; Sandt, E. van de; Ottens, M.
Source Biotechnology and Bioengineering 109 (2012)12. - ISSN 0006-3592 - p. 3070 - 3083.
DOI http://dx.doi.org/10.1002/bit.24576
Department(s) Bioprocess Engineering
VLAG
Publication type Refereed Article in a scientific journal
Publication year 2012
Keyword(s) ion-exchange chromatography - expert-systems - separations - adsorption - optimization - displacers - selection - design
Abstract A multi-dimensional fractionation and characterization scheme was developed for fast acquisition of the relevant molecular properties for protein separation from crude biological feedstocks by ion-exchange chromatography (IEX), hydrophobic interaction chromatography (HIC) and size-exclusion chromatography (SEC). In this approach, the linear IEX isotherm parameters were estimated from multiple linear salt-gradient IEX data, while. the nonlinear IEX parameters as well as the HIC isotherm parameters were obtained by the inverse method under column overloading conditions. Collected chromatographic fractions were analyzed by gel electrophoresis for estimation of molecular mass, followed by mass spectrometry for protein identification. The usefulness of the generated molecular properties data for rational decision-making during downstream process development was equally demonstrated. Monoclonal antibody (mAb) purification from crude hybridoma cell culture supernatant (HCCS) was used as case study. The obtained chromatographic parameters only apply to the employed stationary phases and operating conditions, hence prior HTS of different chromatographic resins and mobile phase conditions is still a prerequisite. Nevertheless, it provides a quick, knowledge-based approach for rationally synthesizing purification cascades prior to more detailed process optimization and evaluation.
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