Separation of biomolecules using Open-Loop Non-Isocratic Simulated moving bed (SMB) technology

Abstract

In biotechnological industries, downstream processing is a bottleneck in the complete production process. There is a wish to reduce the costs and therefore one tries to operate separation technology in continuous mode.

Conventional SMB is a continuous countercurrent separation process, which is based on mathematical modeling and simulation studies for separation of binary mixtures of components. Its application has been limited due to the complexity of the process and involvement of mathematical modeling and simulation. In this thesis, open loop non-isocratic SMB (OLNI-SMB) technology is introduced and developed through an optimized single batch run. Using this technology, proteins from complex multi-component crude samples can be purified. Real industrial waste effluent from food industries such as whey form cheese- and potato fruit juice from starch-producing industries were used for capturing and concentrating proteins through ion exchanger leading to a final recovery of 50% and 80% respectively. It was also used for purification of single target molecules, such as cloned β-glucosidase with a his6-tag that was purified to a purification factor of 15 and a purity of 91%. In this technology, fine-tuning of the separation process is possible by easy and quick change in configuration affecting productivity and the target concentration of the final product as shown by moving a column from loading zone to elution zone resulting in increase of recovery of 20% and a 2-fold increase in protein concentration, respectively. When compared to an equivalent batch separation process, OLNI- SMB technology has shown clear advantage in terms of increase in productivity at least by 20%, 3-fold increase in target concentration and increase in recovery to at least 1.4 times. Another advantage is the reduction in buffer consumption due to continuous operation to 60% or greater in different cases studied. The possibility of further reducing it by recycling the process effluent as a part of chromatographic buffer system as compared from one SMB configuration to another leads to total reduction of 76%. The developed technology can be applied not just for separation of proteins but other areas of separation such as purification of target surfactant from its reaction mixture which led to increase in productivity of 1.5 fold, reduction in buffer by 82% and increase in product concentration by 3 times.

Abstract (Swedish)

Popular Abstract in English

In biotechnological industries, downstream processing is a bottleneck in the complete production process. There is a wish to reduce the costs and therefore one tries to operate separation technology in continuous mode.

Conventional SMB is a continuous countercurrent separation process, which is based on mathematical modeling and simulation studies for separation of binary mixtures of components. Its application has been limited due to the complexity of the process and involvement of mathematical modeling and simulation. In this thesis, open loop non-isocratic SMB (OLNI-SMB) technology is introduced and developed through an optimized single batch run. Using this technology, proteins from complex multi-component crude samples can be purified. Real industrial waste effluent from food industries such as whey form cheese- and potato fruit juice from starch-producing industries were used for capturing and concentrating proteins through ion exchanger leading to a final recovery of 50% and 80% respectively. It was also used for purification of single target molecules, such as cloned β-glucosidase with a his6-tag that was purified to a purification factor of 15 and a purity of 91%. In this technology, fine-tuning of the separation process is possible by easy and quick change in configuration affecting productivity and the target concentration of the final product as shown by moving a column from loading zone to elution zone resulting in increase of recovery of 20% and a 2-fold increase in protein concentration, respectively. When compared to an equivalent batch separation process, OLNI- SMB technology has shown clear advantage in terms of increase in productivity at least by 20%, 3-fold increase in target concentration and increase in recovery to at least 1.4 times. Another advantage is the reduction in buffer consumption due to continuous operation to 60% or greater in different cases studied. The possibility of further reducing it by recycling the process effluent as a part of chromatographic buffer system as compared from one SMB configuration to another leads to total reduction of 76%. The developed technology can be applied not just for separation of proteins but other areas of separation such as purification of target surfactant from its reaction mixture which led to increase in productivity of 1.5 fold, reduction in buffer by 82% and increase in product concentration by 3 times.

• author: Sahoo, Deepti
• supervisor: Bo Mattiasson
• opponent: John Hobley, Timothy, Associate Professor
• organization: Biotechnology
• publishing date: 2012
• type: Thesis
• publication status: published
• subject: Industrial Biotechnology
• keywords: Open-loop Non-Isocratic SMB, Simulated moving bed, True moving bed Chromatography, continuous separation, protein purification, batch chromatography
• defense location: Lecture hall C, Getingevägen 60, Lund University, Faculty of Engineering
• defense date: 2012-11-30 10:30:00
• ISBN: 978-91-89627-88-8
• language: English
• LU publication?: yes
• id: ce1bd894-9ff4-4143-8822-312f81344d41 (old id 3164350)
• date added to LUP: 2016-04-04 12:59:19
• date last changed: 2018-11-21 21:11:40