Evaluation of a new non-alpha il-2 mutein purification scheme.

The elution by imidazole is used in the purification process of non-alpha IL-2 mutein, specifically in the immobilized metal chelates affinity chromatography step (IMAC). In order to demonstrate that a new elution process is possible using cheaper solutions, changes were made in the former capture step. These changes consist on separate the size exclusion chromatography and the controlled oxidation stage which are currently performed together, as well as changing the elution solution. To achieve this, a 22 factorial design of experiments was performed to evaluate the effect of pH and molar concentration on recovery, purity, and particle diameter. The best results were obtained with the solution of sodium acetate 20 mmol L-1 pH 4 being similar to those obtained in the industry nowadays: 29 % recovery, 90 % of purity, and particle diameter 18,40 nm. The first contribution of this paper is to demonstrate the feasibility of using the new elution methodology. Moreover, by using the academic software SuperPro Designer, the actual capture step of the purification process (elution by Imidazole) was simulated, validated, and verified. To demonstrate the relationship between costs and benefits, the CH SFF matrix gel volume was incremented. This change increased costs at this stage. However, the productivity obtained grew to 109 mg h-1, 2.78 times greater than the current productivity. Therefore, the gross net increased by 879 325, 00 $. This result constitutes the second contribution of this paper. [ABSTRACT FROM AUTHOR]