Magnetically stabilised fluidised bed adsorption: practical benefit of uncoupling bed expansion from fluid velocities in the purification of a recombinant protein from Escherichia coli
✍ Scribed by Zhanren Zhang; Deirdre A O'Sullivan; Andrew Lyddiatt
- Publisher
- Wiley (John Wiley & Sons)
- Year
- 1999
- Tongue
- English
- Weight
- 156 KB
- Volume
- 74
- Category
- Article
- ISSN
- 0268-2575
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✦ Synopsis
A composite magnetite-agarose adsorbent has been fabricated in a bead emulsiücation process and, following derivatisation with a peptide, exploited in the direct, one-step puriücation to near molecular homogenity of an anti-MUC1 diabody fragment (dbFv) expressed in the periplasm of recombinant Escherichia coli. Variation of performance with temperature and feedstock concentration was attributed respectively to conformational variation of the immobilised ligand and diþ erent kinetic driving forces at adsorption. The low particle density (1.12 g cm-3) resulting from the low mass percentage of introduced magnetite required that adsorption be operated in a magnetically stabilised ýuidised bed (MSFB) contactor. The contactor achieved direct adsorption of product from EDTAtreated E coli whole broth while operating under conditions of bed expansion uncoupled from ýuid velocity and viscosity. The generic applicability of such adsorbents and contactors to the direct biorecovery of proteins from complex feedstock is discussed.