Downstream process intensification by innovative material development

  • Due to the development of industrial biotechnology where the high level of the desired product is a reality now, there is a need for efficient purification process design of these entities. The integration of a few downstream processing steps could be the solution to achieve highly pure bio-molecules in less time and with fewer resources. This thesis is based on intensification and integration of downstream processing via incorporation of innovative materials to the separation process. Innovative 3D megaporous and imprinted materials were synthesized by free radical initiation of vinyl monomers. A rigid three-dimensional mega porous monolithic structure was produced by polymerization of selected monomers in the presence of a highly soluble cross-linking agent. The megaporous materials bottleneck is a lower binding capacity of bio-molecules, which renders them useless at the industrial level. Particle embedding, direct chemical synthesis or grafting (radiation-induced and chemical induced) was incorporated to enhance the binding capacity. The material was now able to integrate capture and early purification steps and thus was able to be used for process intensification and integration at an industrial level. Various functionalities were incorporated into the megaporous material after grafting of glycidyl methacrylate to the pore surface. In ion-exchange mode, the dynamic binding capacity for cation-exchanger reached 353 mg per g (10% breakthrough) and 507 mg per g (50% breakthrough) and for anion-exchanger was 58 mg per g. In the form of a metal affinity material, it was able to purify 12 mg per g of His-tagged protein, which shows a single band on SDS-PAGE that confirms the purity and integrity of the target protein. The affinity material was created by covalent immobilization of short peptide on the megaporous backbone, which has the affinity for Immuno globulin. The material was compatible with the routine affinity material possessing protein A ligand, which was synthesized and fabricated in the same way of megaporous material. Finally, the megaporous material was used for enzyme immobilization. Lysozyme supported on a grafted body showed extensive clarification activity against a Micrococcus lysodekticus suspension in the flow-through mode. This experiment proved the materials ability to allow the passage of whole cells without clogging the packed bed. Both protein capture and biocatalysis applications are conceivable with the 3D-megaporous materials. Overall, the system was competent to be used efficiently under chromatographic conditions and thus could be efficiently used for process intensification. A process was designed for the direct capture of low molecular weight bio-molecule (carminic acid, CA); the same adsorbent was not feasible to be used for purification of low molecular weight substances. Therefore, an innovative rigid 3-D beaded molecularly imprinted megaporous material was synthesized by radical polymerization of monomer and cross-linker for process intensification of low molecular weight bio-molecules. The synthesis strategy developed here was a novel method based on the non - covalent approach, which resulted in highly specific particles possessing the high binding capacity. The observed data (surface area, charge, imprinting factor, selectivity factor, maximum capacity and maximum available binding sites) indicates that the imprinted adsorbent could be conveniently utilized for the recovery of CA from cochineal extract in the finite bath mode of operation. The material was able specifically to capture CA from clarified extract in a single step. Mass transfer limitation is the major problem that renders the use of adsorbents from industrial level. In the materials produced here, the mass transfer resistance was evaluated and it was observed that most of the resistances come from the pore diffusion. The increase in the bulk movements was enough to overcome the resistance.

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Publishing Institution:IRC-Library, Information Resource Center der Jacobs University Bremen
Granting Institution:Jacobs Univ.
Author:Noor Shad Bibi
Referee:Marcelo Fernandez Lahore, Thomas Nugent, Herbert Hugl
Advisor:Marcelo Fernandez Lahore
Persistent Identifier (URN):urn:nbn:de:101:1-201305294727
Document Type:PhD Thesis
Language:English
Date of Successful Oral Defense:2012/05/29
Year of Completion:2012
Date of First Publication:2012/06/22
PhD Degree:Biochemical Engineering
School:SES School of Engineering and Science
Library of Congress Classification:T Technology / TP Chemical technology / TP248.13-248.65 Biotechnology / TP248.3 Biochemical engineering. Bioprocess engineering
Call No:Thesis 2012/13

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