title: Expanded bed adsorption as a primary recovery step for the isolation of the insulin precursor MI3 process development and scale up.
abstract: 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.
authors.affiliation: Institute of Enzyme Technology, Heinrich-Heine University Düsseldorf, 52426 Jülich, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Brixius
authors.forename: Inger
authors.initials: I
authors.lastname: Mollerup
authors.forename: Ole Elvang
authors.initials: OE
authors.lastname: Jensen
authors.forename: Markus
authors.initials: M
authors.lastname: Halfar
authors.forename: Jörg
authors.initials: J
authors.lastname: Thömmes
authors.forename: Maria-Regina
authors.initials: MR
authors.lastname: Kula
mesh_terms: D000327:Adsorption; D002853:Chromatography, Liquid / Q000379:methods; D006801:Humans; D011384:Proinsulin / Q000096:biosynthesis / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000254:growth & development*
chemical_list: D011994:Recombinant Proteins; D011384:Proinsulin