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