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Comparison of affinity membrane adsorbers for the purification of recombinant human erythropoietin (rhEPO)
Author(s) -
Mellado Maria Candida M,
Curbelo David,
Nobrega Ronaldo,
Castilho Leda R
Publication year - 2007
Publication title -
journal of chemical technology and biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.64
H-Index - 117
eISSN - 1097-4660
pISSN - 0268-2575
DOI - 10.1002/jctb.1720
Subject(s) - membrane , chemistry , wheat germ agglutinin , concanavalin a , monoclonal antibody , affinity chromatography , chromatography , recombinant dna , erythropoietin , biochemistry , lectin , antibody , biology , in vitro , enzyme , gene , immunology , endocrinology
In this work affinity membrane adsorbers were investigated for the chromatographic purification of recombinant human erythropoietin (rhEPO) produced in mammalian cells. Cibacron Blue (CB), IDA‐Cu +2 , wheat germ agglutinin (WGA), concanavalin A (ConA) and an anti‐EPO monoclonal antibody (MAb) were tested as affinity ligands, attached to microporous Sartobind ® membranes. In experiments carried out with cell culture supernatant, the best results were obtained with Sartobind–CB, Sartobind–WGA and Sartobind–MAb membranes. The thermodynamic parameters were determined by adsorption isotherms of rhEPO onto the membranes. Sartobind–ConA presented the lowest affinity for rhEPO, as evidenced by a lower association constant. For Sartobind–CB, Sartobind–IDA‐Cu +2 and Sartobind–MAb K A was in the order of 10 5 L mol −1 , whereas for Sartobind–WGA it was 10 6 L mol −1 . Sartobind–CB eluates were also investigated by RP‐HPLC. The purity level achieved in this one‐step purification strategy was 55%, indicating that the Sartobind–CB membrane is a promising affinity membrane for rhEPO purification. Copyright © 2007 Society of Chemical Industry