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Primary Recovery of Biologically Active Compounds Using Macroporous Monolithic Hydrogels

Hanora, Amro LU (2005)
Abstract
The completion of sequencing of human and other mammalian genomes opens a new era for drug development. New approaches have been proposed for tackling diseases such as development of small molecule therapies based on understanding of human genome and proteome, new proteinous based therapies, where the therapeutic protein can inhibit or modify a certain metabolic pathway or DNA-based therapies delivering a gene into the cell (e.g., cancer cells) to correct or inhibit the imperfect gene function. The possibility of production of novel therapeutics (small, proteinous and DNA based substances) in biotechnological processes utilising mammalian cells or microorganisms such as bacteria or yeast, presents new challenges for the separation... (More)
The completion of sequencing of human and other mammalian genomes opens a new era for drug development. New approaches have been proposed for tackling diseases such as development of small molecule therapies based on understanding of human genome and proteome, new proteinous based therapies, where the therapeutic protein can inhibit or modify a certain metabolic pathway or DNA-based therapies delivering a gene into the cell (e.g., cancer cells) to correct or inhibit the imperfect gene function. The possibility of production of novel therapeutics (small, proteinous and DNA based substances) in biotechnological processes utilising mammalian cells or microorganisms such as bacteria or yeast, presents new challenges for the separation processes. Moreover, new methods for high throughput screening are needed to identify potentially important lead molecules within the large molecular libraries. Indeed, designing downstream process (DSP) of biologically active compounds depends on the nature and intended use of such compounds. Commodity compounds that are produced in tons need fast and cheap purification methods. However, high molecular weight molecules intended for pharmaceutical applications, need careful design of DSP to meet the legislator demands such as removal of contaminants and toxic substances. Two examples were demonstrated for the design of DSP purification process for small and large molecular weight compounds. Lactic acid was purified from cell broth using composite ion exchanger in expanded bed chromatographic format. Plasmid DNA was captured directly from cell lysate using monolithic macroporous hydrogel, so called cryogel, grafted with different polycations. Using cryogels with coupled specific ligands, bacterial endotoxin was successfully removed from protein solutions and captured from disposed cell homogenate. Fast parallel screening of complex samples containing target substance was demonstrated using affinity minicryogel columns in 96-well format plate. (Less)
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author
supervisor
opponent
  • Associate Professor Hobley, Timothy John, BioCentrum, Technical University of Denmark (TUD), Denmark.
organization
publishing date
type
Thesis
publication status
published
subject
keywords
Downstream processing, macroporous, Biologically active compounds, Biotechnology, Läkemedelsteknik och relaterad teknik, Pharmaceutical and related technologies, Polymerteknik, biopolymers, Biokemisk teknik, Polymer technology, Makromolekylär kemi, Biochemical technology, Affinity chromatography, Ion exchange chromatography, Expanded bed adsorption chromatography., Macromolecular chemistry, High through put screening, lactica acids, Endotoxin, Plasmid DNA, Cryogel columns, Bioteknik
pages
100 pages
publisher
Biotechnology, Lund University
defense location
Lecture hall A, at the Center for Chemistry and Chemical Engineering, Sölvegatan 39, Lund Institute of Technology
defense date
2005-11-09 10:30:00
external identifiers
  • other:ISRN: LUTKDH/TKBT--05/1088--SE
ISBN
91-89627-35-0
language
English
LU publication?
yes
additional info
Irina Savina, Amro Hanora, Fatima Plieva, Igor Galaev, Bo Mattiasson and Vladimir Lozinsky. 2004. Cryostructuration of Polymer Systems. XXIV. Poly(vinyl alcohol) Cryogels Filled with Particles of a Strong Anion Exchanger: Properties of the Composite Materials and Potential Applications Journal of Applied Polymer Science, vol 95 pp 529-538. Biotechnology department, Chemical center, Lund University
Amro Hanora, Florent Bernaudat, Fatima Plieva, Maria Dainiak, Leif Bülow, Igor Galaev and Bo Mattiasson. 2005. Screening of peptide affinity tags using immobilised metal affinity chromatography in 96-well plate format Journal of Chromatography A, vol 1087 pp 38-“44. Biotechnology department, Chemical Center, Lund University
Amro Hanora, Fatima Plieva, Martin Hedström, Igor Galaev and Bo Mattiasson. 2005. Capture of bacterial endotoxins using a supermacroporous monolithic matrix with immobilized polyethyleneimine,lysozyme or polymyxin B Journal of Biotechnology, vol 118 pp 421-433. Biotechnology department, Chemical Center, Lund University
Amro Hanora, Irina Savina, Fatima Plieva, Vladimir Izumrudov, Bo Mattiasson and Igor Galaev. 2005. Direct capture of bacterial plasmid DNA from non-clarified cell lysate using monolith columns from macroporous gel grafted with polycations Journal of biotechnology, Biotechnology department, Chemical Center, Lund University (submitted)
id
1ec14e3a-918d-4f70-a965-e6600db2e5ad (old id 545390)
date added to LUP
2016-04-04 09:58:36
date last changed
2018-11-21 20:55:57
@phdthesis{1ec14e3a-918d-4f70-a965-e6600db2e5ad,
  abstract     = {{The completion of sequencing of human and other mammalian genomes opens a new era for drug development. New approaches have been proposed for tackling diseases such as development of small molecule therapies based on understanding of human genome and proteome, new proteinous based therapies, where the therapeutic protein can inhibit or modify a certain metabolic pathway or DNA-based therapies delivering a gene into the cell (e.g., cancer cells) to correct or inhibit the imperfect gene function. The possibility of production of novel therapeutics (small, proteinous and DNA based substances) in biotechnological processes utilising mammalian cells or microorganisms such as bacteria or yeast, presents new challenges for the separation processes. Moreover, new methods for high throughput screening are needed to identify potentially important lead molecules within the large molecular libraries. Indeed, designing downstream process (DSP) of biologically active compounds depends on the nature and intended use of such compounds. Commodity compounds that are produced in tons need fast and cheap purification methods. However, high molecular weight molecules intended for pharmaceutical applications, need careful design of DSP to meet the legislator demands such as removal of contaminants and toxic substances. Two examples were demonstrated for the design of DSP purification process for small and large molecular weight compounds. Lactic acid was purified from cell broth using composite ion exchanger in expanded bed chromatographic format. Plasmid DNA was captured directly from cell lysate using monolithic macroporous hydrogel, so called cryogel, grafted with different polycations. Using cryogels with coupled specific ligands, bacterial endotoxin was successfully removed from protein solutions and captured from disposed cell homogenate. Fast parallel screening of complex samples containing target substance was demonstrated using affinity minicryogel columns in 96-well format plate.}},
  author       = {{Hanora, Amro}},
  isbn         = {{91-89627-35-0}},
  keywords     = {{Downstream processing; macroporous; Biologically active compounds; Biotechnology; Läkemedelsteknik och relaterad teknik; Pharmaceutical and related technologies; Polymerteknik; biopolymers; Biokemisk teknik; Polymer technology; Makromolekylär kemi; Biochemical technology; Affinity chromatography; Ion exchange chromatography; Expanded bed adsorption chromatography.; Macromolecular chemistry; High through put screening; lactica acids; Endotoxin; Plasmid DNA; Cryogel columns; Bioteknik}},
  language     = {{eng}},
  publisher    = {{Biotechnology, Lund University}},
  school       = {{Lund University}},
  title        = {{Primary Recovery of Biologically Active Compounds Using Macroporous Monolithic Hydrogels}},
  url          = {{https://lup.lub.lu.se/search/files/5430783/545395.pdf}},
  year         = {{2005}},
}