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Faster isolation of recombinant factor VIII SQ, with a superporous agarose matrix

Pålsson, Eva ; Smeds, Anna-Lisa ; Peterson, Anna and Larsson, Per-Olof LU (1999) In Journal of chromatography. A 840(1). p.39-50
Abstract
A superporous agarose matrix was compared with a corresponding homogenous matrix in the isolation of recombinant factor VIII SQ (r-VIII SQ) by immunoaffinity chromatography. As a reference, the commercially available Sepharose FastFlow, used for a similar purification in the industry, was also evaluated. Breakthrough curves are described for flows between 50 and 400 cm/h with pre-purified r-VIII SQ and with cell culture broth. The superporous gel gave the best performance and a 1000-fold purification was obtained in a one-step procedure. The superporous matrix made it possible to increase the throughput about four-fold, presumably due to its better mass transfer properties. The importance of the ligand distribution profile is discussed... (More)
A superporous agarose matrix was compared with a corresponding homogenous matrix in the isolation of recombinant factor VIII SQ (r-VIII SQ) by immunoaffinity chromatography. As a reference, the commercially available Sepharose FastFlow, used for a similar purification in the industry, was also evaluated. Breakthrough curves are described for flows between 50 and 400 cm/h with pre-purified r-VIII SQ and with cell culture broth. The superporous gel gave the best performance and a 1000-fold purification was obtained in a one-step procedure. The superporous matrix made it possible to increase the throughput about four-fold, presumably due to its better mass transfer properties. The importance of the ligand distribution profile is discussed based upon immunofluorescence microscopy data. (Less)
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author
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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Immunoaffinity chromatography, Comparison with commercial adsorbent
in
Journal of chromatography. A
volume
840
issue
1
pages
39 - 50
publisher
Elsevier
external identifiers
  • scopus:0343448992
ISSN
0021-9673
DOI
10.1016/S0021-9673(99)00209-5
language
English
LU publication?
yes
id
09fe7e2f-1943-42b6-b9c3-73df80b2de6d
date added to LUP
2024-05-17 21:52:33
date last changed
2024-05-30 11:32:04
@article{09fe7e2f-1943-42b6-b9c3-73df80b2de6d,
  abstract     = {{A superporous agarose matrix was compared with a corresponding homogenous matrix in the isolation of recombinant factor VIII SQ (r-VIII SQ) by immunoaffinity chromatography. As a reference, the commercially available Sepharose FastFlow, used for a similar purification in the industry, was also evaluated. Breakthrough curves are described for flows between 50 and 400 cm/h with pre-purified r-VIII SQ and with cell culture broth. The superporous gel gave the best performance and a 1000-fold purification was obtained in a one-step procedure. The superporous matrix made it possible to increase the throughput about four-fold, presumably due to its better mass transfer properties. The importance of the ligand distribution profile is discussed based upon immunofluorescence microscopy data.}},
  author       = {{Pålsson, Eva and Smeds, Anna-Lisa and Peterson, Anna and Larsson, Per-Olof}},
  issn         = {{0021-9673}},
  keywords     = {{Immunoaffinity chromatography; Comparison with commercial adsorbent}},
  language     = {{eng}},
  number       = {{1}},
  pages        = {{39--50}},
  publisher    = {{Elsevier}},
  series       = {{Journal of chromatography. A}},
  title        = {{Faster isolation of recombinant factor VIII SQ, with a superporous agarose matrix}},
  url          = {{http://dx.doi.org/10.1016/S0021-9673(99)00209-5}},
  doi          = {{10.1016/S0021-9673(99)00209-5}},
  volume       = {{840}},
  year         = {{1999}},
}