Improved Recovery of Erythropoietin and Darbepoetin from Equine Plasma by the Application of a Wheat Germ Agglutinin Mediated Pre-Extraction Prior to Immunoaffinity Chromatography
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Affiliation(s)
The Singapore Turf Club Laboratory, 1 Turf Club Ave, Singapore, Singapore.
The Singapore Turf Club Laboratory, 1 Turf Club Ave, Singapore, Singapore.
The Singapore Turf Club Laboratory, 1 Turf Club Ave, Singapore, Singapore.
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.
KEYWORDS
Erythropoietin, Darbepoetin Alfa, Immunoaffinity Extraction, Lectin Affinity Extraction, Wheat Germ Agglutinin, Horseracing
Cite this paper
References
Stanley,
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Agglutinin Mediated Pre-Extraction Prior to Immunoaffinity
Chromatography. Advances in Bioscience and Biotechnology, 5, 651-660. doi: 10.4236/abb.2014.57077.
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