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Expanded bed adsorption as a primary recovery step for the isolation of the insulin precursor MI3 process development and scale up
Author(s) -
Brixius Peter,
Mollerup Inger,
Jensen Ole Elvang,
Halfar Markus,
Thömmes Jörg,
Kula MariaRegina
Publication year - 2005
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20531
Subject(s) - expanded bed adsorption , adsorption , chromatography , isoelectric point , residence time distribution , chemistry , scale up , yield (engineering) , process (computing) , chemical engineering , materials science , computer science , organic chemistry , engineering , mineralogy , enzyme , inclusion (mineral) , physics , classical mechanics , metallurgy , operating system
Expanded bed adsorption (EBA) was evaluated for the isolation of the human insulin precursor MI3, expressed and secreted by the yeast Saccharomyces cerevisiae . The isoelectric point of the insulin precursor (pH 5.3) makes cation exchange a prime candidate for direct adsorption. In order to find a suitable window of operation for the process the adsorption equilibrium was analysed in a wide range of operating conditions (pH and conductivity) and for three different stationary phases. The same array of operating conditions was examined with regard to stable fluidisation of the adsorbents in S. cerevisiae suspensions. Interactions of the yeast with the fluidised stationary phase were investigated by a pulse response technique and the hydrodynamics of the fluidised bed under process conditions by residence time distribution analysis. The case study demonstrates that by parallel examination of product binding and fluidisation quality a window of operation can be found. Analysis of the binding kinetics by breakthrough experiments and modelling led to the definition of a set of operating conditions, which yield a compromise between optimal use of the equilibrium capacity provided by the adsorbent and high throughput required for an industrial separation. After initial experiments on the bench scale the protocol was transferred successfully to pilot scale demonstrating the design of a reliable operation. © 2005 Wiley Periodicals, Inc.