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Determination of diffusion coefficients of proteins in stationary phases by frontal chromatography

Kempe, Henrik LU ; Persson, Patrik LU ; Axelsson, Anders LU ; Nilsson, Bernt LU and Zacchi, Guido LU (2006) In Biotechnology and Bioengineering 93(4). p.656-664
Abstract
A strategy to determine effective diffusion coefficients of proteins in chromatographic gels is presented in this article. An experimental methodology based on frontal liquid chromatography was combined with a numerical methodology based on a mathematical model describing the chromatographic process including the extra-column dispersion, the dispersion due to the packed bed, the external mass transfer from the bulk phase to the stationary phase, and the diffusive transport within the stationary phase. The methodology has several advantages compared to previously reported methods to determine diffusion coefficients in that no other equipment than an HPLC is required, any class of stationary phases can be investigated as long as the... (More)
A strategy to determine effective diffusion coefficients of proteins in chromatographic gels is presented in this article. An experimental methodology based on frontal liquid chromatography was combined with a numerical methodology based on a mathematical model describing the chromatographic process including the extra-column dispersion, the dispersion due to the packed bed, the external mass transfer from the bulk phase to the stationary phase, and the diffusive transport within the stationary phase. The methodology has several advantages compared to previously reported methods to determine diffusion coefficients in that no other equipment than an HPLC is required, any class of stationary phases can be investigated as long as the experiments are performed under non-binding conditions, and no modification, e.g., moulding of slabs or membranes, to the stationary phase is required. To show the applicability of the methodology, the effective diffusion coefficients of lysozyme, bovine serum albumin, and immunoglobulin gamma in Sepharose (TM) CL-4B were determined and shown to be comparable with those determined with other methods. (c) 2005 Wiley Periodicals, Inc. (Less)
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author
; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
effective diffusion coefficient, diffusion, chromatography, frontal, protein separation, downstream processing
in
Biotechnology and Bioengineering
volume
93
issue
4
pages
656 - 664
publisher
John Wiley and Sons
external identifiers
  • wos:000235630600006
  • pmid:16372360
  • scopus:33644913140
ISSN
1097-0290
DOI
10.1002/bit.20738
language
English
LU publication?
yes
id
d896b7e7-c80a-4159-9436-fa5b5f171529 (old id 417230)
date added to LUP
2016-04-01 11:36:32
date last changed
2021-06-30 05:03:48
@article{d896b7e7-c80a-4159-9436-fa5b5f171529,
  abstract     = {A strategy to determine effective diffusion coefficients of proteins in chromatographic gels is presented in this article. An experimental methodology based on frontal liquid chromatography was combined with a numerical methodology based on a mathematical model describing the chromatographic process including the extra-column dispersion, the dispersion due to the packed bed, the external mass transfer from the bulk phase to the stationary phase, and the diffusive transport within the stationary phase. The methodology has several advantages compared to previously reported methods to determine diffusion coefficients in that no other equipment than an HPLC is required, any class of stationary phases can be investigated as long as the experiments are performed under non-binding conditions, and no modification, e.g., moulding of slabs or membranes, to the stationary phase is required. To show the applicability of the methodology, the effective diffusion coefficients of lysozyme, bovine serum albumin, and immunoglobulin gamma in Sepharose (TM) CL-4B were determined and shown to be comparable with those determined with other methods. (c) 2005 Wiley Periodicals, Inc.},
  author       = {Kempe, Henrik and Persson, Patrik and Axelsson, Anders and Nilsson, Bernt and Zacchi, Guido},
  issn         = {1097-0290},
  language     = {eng},
  number       = {4},
  pages        = {656--664},
  publisher    = {John Wiley and Sons},
  series       = {Biotechnology and Bioengineering},
  title        = {Determination of diffusion coefficients of proteins in stationary phases by frontal chromatography},
  url          = {http://dx.doi.org/10.1002/bit.20738},
  doi          = {10.1002/bit.20738},
  volume       = {93},
  year         = {2006},
}