Beckley K. Nfor1, Tangir Ahamed1, Marcel Ottens1, Emile J. A. X. van de Sandt2, Michel Eppink3, Gijs W. K. van Dedem1, and Luuk A. M. van der Wielen1. (1) Department of Biotechnology, Faculty of Applied Sciences, Delft University of Technology, Julianalaan 67, Delft, 2628 BC, Netherlands, (2) DSM Anti-infectives BV, P.O.Box 425, Delft, 2600 AK, Netherlands, (3) DSP, Diosynth Biotechnology Europe, Oss, Netherlands
Protein bioseparation processes can generally be subdivided into protein recovery for the isolation of the target protein from the fermentation broth and its concentration in the cell-free solution, followed by purification of the target protein to sufficiently high purity. While the selection and sequencing of protein recovery operations is rather straightforward, this is not the case with protein purification due to the fact that several feasible combinations of protein purification techniques may be possible, from which the best one has to be selected. Systematic synthesis and design of protein purification processes is currently limited by two main factors, namely: (1) the general lack of the relevant thermodynamic properties of proteins and (2) the lack of a rational approach for selecting and sequencing protein purification techniques. Consequently, the selection and sequencing of protein purification techniques is mostly carried out qualitatively, based on substantial experience and heuristics, often resulting in overall suboptimal downstream processing. In our recent study [1], the first problem above was addressed through the development of generalized predictive thermodynamic models of macromolecules that are applicable to a number of bioseparation techniques (SEC, HIC, RPC, ATPS, crystallization and precipitation). Here, we address the second problem above by innovating protein purification process synthesis via the use of quantitative elements (mass balances, thermodynamic models, design equations and performance indicators such as recovery yield, purity, productivity and process economics) as well as a High Throughput Experimentation (HTE) methodology for quickly acquiring the necessary experimental data.
Reference: [1] T. Ahamed, M. Ottens, B. K. Nfor, G. W. K. van Dedem, L. A. M. van der Wielen. Fluid Phase Equilibria, 241, 268-282 (2006).
Acknowledgement: This project is financially supported by the Netherlands Ministry of Economic Affairs and the B-Basic partner organizations (www.b-basic.nl) through B-Basic, a public-private NWO-ACTS programme (ACTS = Advanced Chemical Technologies for Sustainability).