TITLE:
Improved Recovery of Erythropoietin and Darbepoetin from Equine Plasma by the Application of a Wheat Germ Agglutinin Mediated Pre-Extraction Prior to Immunoaffinity Chromatography
AUTHORS:
Shawn M. R. Stanley, Danny Chua
KEYWORDS:
Erythropoietin, Darbepoetin Alfa, Immunoaffinity Extraction, Lectin Affinity Extraction, Wheat Germ Agglutinin, Horseracing
JOURNAL NAME:
Advances in Bioscience and Biotechnology,
Vol.5 No.7,
June
27,
2014
ABSTRACT:
We describe a two-step method that uses wheat germ agglutinin immobilized
on Sepharose gel followed by immunoaffinity chromatography (IAC) to extract
recombinant human erythropoietin and Darbepoetin from equine plasma. Lectin
affinity chromatography was shown to be an effective approach for isolating
these epoetins from plasma and in combination with IAC; this method gave
superior recovery when compared to the use of the latter technique alone.
Moreover, due to the ease with which it can be scaled up, it is particularly
well suited for pre-concentrating larger volumes of samples prior to IAC and
this provides a facile way of improving the overall sensitivity with which
these foreign proteins can be detected in equine plasma.