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Department of Pharmacology

 
Author(s): 
El Khoury, G, Wang, Y, Wang, D, Jacob, SI, Lowe, CR
Abstract: 

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 ∼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.

Publication ID: 
542242
Published date: 
November 2013
Publication source: 
pubmed
Publication type: 
Journal articles
Journal name: 
Biotechnol Bioeng
Publication volume: 
110
Publisher: 
Parent title: 
Edition: 
Publication number: