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Magnetically stabilised fluidised bed adsorption: practical benefit of uncoupling bed expansion from fluid velocities in the purification of a recombinant protein from Escherichia coli
Author(s) -
Zhang Zhanren,
O'Sullivan Deirdre A,
Lyddiatt Andrew
Publication year - 1999
Publication title -
journal of chemical technology and biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.64
H-Index - 117
eISSN - 1097-4660
pISSN - 0268-2575
DOI - 10.1002/(sici)1097-4660(199903)74:3<270::aid-jctb24>3.0.co;2-x
Subject(s) - expanded bed adsorption , adsorption , chromatography , chemistry , contactor , agarose , raw material , chemical engineering , magnetite , materials science , organic chemistry , thermodynamics , metallurgy , power (physics) , physics , engineering
A composite magnetite–agarose adsorbent has been fabricated in a bead emulsification process and, following derivatisation with a peptide, exploited in the direct, one‐step purification 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 different 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 fluidised bed (MSFB) contactor. The contactor achieved direct adsorption of product from EDTA‐treated E coli whole broth while operating under conditions of bed expansion uncoupled from fluid velocity and viscosity. The generic applicability of such adsorbents and contactors to the direct biorecovery of proteins from complex feedstock is discussed. © 1999 Society of Chemical Industry