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.

Experimental characterization of next‐generation expanded‐bed adsorbents for capture of a recombinant protein expressed in high‐cell‐density yeast fermentation

“…This result indicates that a change to the steps between load and elution might result in an improved yield. These results compare favorably to the 75% yield obtained by Chen and Sun in recovering BSA from a 6% yeast solution with DEAE resin, and the 89% yield obtained by Thommes and coworkers in recovering an Insulin precursor molecule yeast broth using the cationic exchanger Streamline® SP.…”

Expanded Beds: A Process Solution for Adsorptive Separations in Waste-Water Treatment

“…As it enters the human body, damages the protein, lipids and DNA by generating reactive free oxygen species [1]. The excessive intake of copper may also result in schizophrenia and the Alzheimer kind of diseases [3]- [5]. Copper has a similar adverse effect on marine ecosystem, damaging aquatic life [6]- [8].…”

Screen-less expanded bed column: new approach for the recovery and purification of a malaria transmission blocking vaccine candidate from Pichia pastoris

“…In addition, this configuration is sensitive to air bubbles that can cause turbulence, adversely affecting column stability as well as protein binding, especially at high biomass concentrations. To prevent collapse of the column, careful evaluation of biomass concentrations and flow rates must be done before the column can be operated (Brixius et al 2005;Chang and Chase 1996;Fernandez-Lahore et al 1999;Lin et al 2004). In addition, careful monitoring of the column's performance and operating parameters must be conducted while it is in use.…”