Design, Synthesis, and Assessment of a De Novo Affinity Adsorbent for the Purification of Recombinant Human Erythropoietin

作者：El Khoury Graziella^*; Wang Yian; Wang Di; Jacob Shaleem I; Lowe Christopher R

来源：Biotechnology and Bioengineering, 2013, 110(11): 3063-3069.

DOI：10.1002/bit.24943

• affinity chromatography
• synthetic affinity adsorbent
• purification of rHuEPO
• Ugi multicomponent reaction

This work describes the assessment of a de novo synthetic affinity ligand for recombinant human erythropoietin (rHuEPO), based on the multicomponent Ugi reaction. Four Ugi ligands were designed based on the X-ray crystallographic structure of the complex between human erythropoietin and site 1 of its cell-surface receptor (EPObp)(2); screening of the ligands with pure rHuEPO samples identified a lead ligand (A9C10I8) immobilized on aldehyde-functionalized agarose beads, which was able to bind and elute erythropoietin, as determined by SDS-PAGE and Western blot analyses. Furthermore, small-scale affinity chromatography performed on the immobilized adsorbent showed its ability to isolate rHuEPO from a spiked mammalian cell supernatant with a purity of approximate to 80%, as estimated with gel densitometry. This approach could lead to the development of a cost-effective downstream process for rHuEPO, as an alternative to the current multi-step purification protocols. Biotechnol. Bioeng. 2013;110: 3063-3069.

• 出版日期2013-11