Advanced

Superporous agarose a new material for chromatography

Gustavsson, Per-Erik LU (1998)
Abstract
Agarose, the most commonly employed chromatography base material in industrial purification processes for biomolecules – proteins, has almost ideal properties. The major disadvantage of the agarose support is its poor mechanical strength which excludes its use in high speed separations where small particles are needed in order to achieve improved chromatographic efficiency. This drawback can be reduced by cross-linking the agarose. A different approach, exploited in this work, is to introduce the concept of intraparticle convective flow to be able to use comparatively large agarose beads which tolerate higher flow rates and still achieve high efficiency.



Thus, this thesis describes superporous agarose as a support... (More)
Agarose, the most commonly employed chromatography base material in industrial purification processes for biomolecules – proteins, has almost ideal properties. The major disadvantage of the agarose support is its poor mechanical strength which excludes its use in high speed separations where small particles are needed in order to achieve improved chromatographic efficiency. This drawback can be reduced by cross-linking the agarose. A different approach, exploited in this work, is to introduce the concept of intraparticle convective flow to be able to use comparatively large agarose beads which tolerate higher flow rates and still achieve high efficiency.



Thus, this thesis describes superporous agarose as a support material for chromatography. Superporous agarose beads in sizes varying between 50-500 µm were prepared. Apart from normal diffusion pores, these beads contain large flow pores (about 5-50 µm) in which chromatographed substances are transported by flow to the interior of each bead. This intraparticle convective flow significantly reduces the distance which must be covered by the slow diffusion process giving an improved mass transport.



Superporous agarose beads were derivatized with different functional groups and their performance compared with corresponding homogeneous agarose beads in ion exchange-, affinity- and hydrophobic interaction chromatography separations of proteins. In these chromatographic modes superporous beads performed considerably better than homogeneous beads.



In a separate study, convective fluid velocities inside superporous agarose beads were directly measured for the first time by following the movement of micro particles in a packed bed. The fluid velocity data obtained were compared with and found to agree with theoretically calculated values based on the Kozeny–Carman equation.



Superporous agarose can also be prepared as continuous gels in the form of cast beds, fibres and thick membranes. Applications of such continuous gels are demonstrated and discussed. (Less)
Please use this url to cite or link to this publication:
author
opponent
  • Doc. Hagel, Lars, Amersham Pharmacia Biotech AB, Uppsala, Sweden
organization
publishing date
type
Thesis
publication status
published
subject
keywords
Protein separations, Chromatography, Superporous agarose, Continuous agarose beds, Intraparticle convection, Pore flow, Biochemistry, Metabolism, Biokemi, metabolism, Biotechnology, Bioteknik
pages
154 pages
publisher
Department of Pure and Applied Biochemistry, Lund University
defense location
Lecture hall C at the Center for Chemistry and Chemical Engineering, Sölvegatan 39, Lund
defense date
1998-09-25 13:15
external identifiers
  • Other:ISRN: LUTKDH/TKBK-1045/1-154/1998
ISBN
91-628-3120-8
language
English
LU publication?
yes
id
cfdbd046-c5ac-4b7f-bc03-f11fd0f88fc0 (old id 38954)
date added to LUP
2007-06-21 12:55:57
date last changed
2016-09-19 08:45:15
@phdthesis{cfdbd046-c5ac-4b7f-bc03-f11fd0f88fc0,
  abstract     = {Agarose, the most commonly employed chromatography base material in industrial purification processes for biomolecules – proteins, has almost ideal properties. The major disadvantage of the agarose support is its poor mechanical strength which excludes its use in high speed separations where small particles are needed in order to achieve improved chromatographic efficiency. This drawback can be reduced by cross-linking the agarose. A different approach, exploited in this work, is to introduce the concept of intraparticle convective flow to be able to use comparatively large agarose beads which tolerate higher flow rates and still achieve high efficiency.<br/><br>
<br/><br>
Thus, this thesis describes superporous agarose as a support material for chromatography. Superporous agarose beads in sizes varying between 50-500 µm were prepared. Apart from normal diffusion pores, these beads contain large flow pores (about 5-50 µm) in which chromatographed substances are transported by flow to the interior of each bead. This intraparticle convective flow significantly reduces the distance which must be covered by the slow diffusion process giving an improved mass transport.<br/><br>
<br/><br>
Superporous agarose beads were derivatized with different functional groups and their performance compared with corresponding homogeneous agarose beads in ion exchange-, affinity- and hydrophobic interaction chromatography separations of proteins. In these chromatographic modes superporous beads performed considerably better than homogeneous beads.<br/><br>
<br/><br>
In a separate study, convective fluid velocities inside superporous agarose beads were directly measured for the first time by following the movement of micro particles in a packed bed. The fluid velocity data obtained were compared with and found to agree with theoretically calculated values based on the Kozeny–Carman equation.<br/><br>
<br/><br>
Superporous agarose can also be prepared as continuous gels in the form of cast beds, fibres and thick membranes. Applications of such continuous gels are demonstrated and discussed.},
  author       = {Gustavsson, Per-Erik},
  isbn         = {91-628-3120-8},
  keyword      = {Protein separations,Chromatography,Superporous agarose,Continuous agarose beds,Intraparticle convection,Pore flow,Biochemistry,Metabolism,Biokemi,metabolism,Biotechnology,Bioteknik},
  language     = {eng},
  pages        = {154},
  publisher    = {Department of Pure and Applied Biochemistry, Lund University},
  school       = {Lund University},
  title        = {Superporous agarose a new material for chromatography},
  year         = {1998},
}