Lectin affinity chromatography as a tool to differentiate endogenous and recombinant erythropoietins.
This work exploits the combination of the lectin affinity chromatography (LAC) with an ultra-sensitive immunochromatographic assay to differentiate several types of erythropoietin (EPO). The chromatographic behaviours of different commercial types of recombinant human EPO (rhEPO), EPO analogues (Aranesp) and urine human EPO (uhEPO) from healthy individuals on eight lectin-Sepharose columns, have been worked out. Results show that when using wheat germ agglutinin (WGA)-Sepharose columns, a careful desorption regime starting with very low concentration (2mM) of the competitive sugar N-acetylglucosamine (GlcNAc) makes it possible to efficiently distinguish endogenous EPO from recombinant EPO and EPO analogues.
Franco Fraguas
L
L
Cátedra de Bioquímica, Departamento de Biociencias, Facultad de Quimica, CC 1157, Montevideo, Uruguay. lfranco@fq.edu.uy
Carlsson
J
J
Lönnberg
M
M
D000117:Acetylglucosamine / Q000737:chemistry; D000327:Adsorption; D002846:Chromatography, Affinity / Q000379:methods*; D004921:Erythropoietin / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000652:urine*; D006801:Humans; D007118:Immunoassay / Q000379:methods; D037102:Lectins / Q000737:chemistry*; D011994:Recombinant Proteins; D014909:Wheat Germ Agglutinins / Q000737:chemistry
D037102:Lectins; D011994:Recombinant Proteins; D014909:Wheat Germ Agglutinins; D004921:Erythropoietin; D000117:Acetylglucosamine