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

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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.

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Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Chemical Engineering

Keywords

  • effective diffusion coefficient, diffusion, chromatography, frontal, protein separation, downstream processing
Original languageEnglish
Pages (from-to)656-664
JournalBiotechnology and Bioengineering
Volume93
Issue number4
Publication statusPublished - 2006
Publication categoryResearch
Peer-reviewedYes