Study of the feasibility of using a pellicular anion-exchange column for separation of transferrin isoforms in human serum by HPLC with UV detection.
A method has been optimised for the separation of glycoforms of human serum transferrin, using a high-performance pellicular anion-exchange chromatographic column. The effect of the eluent pH and of the column temperature on the separation of transferrin glycoforms was studied using a standard solution of commercially available human serum transferrin. An HPLC system equipped with an ultraviolet detector was used for the analysis. No immunoassay was used after the anion-exchange chromatographic separation of the glycoforms, in contrast with most currently used methods. The method was applied to the separation and quantification of transferrin glycoforms in serum from a healthy, non-pregnant woman, after saturation of transferrin with iron and further precipitation of lipoproteins. The whole chromatographic run, including re-equilibration of the column, took 35 min.
de la Calle Guntiñas
M B
MB
European Commission, Joint Research Center, Institute for Reference Materials and Measurements, Retieseweg, 2440 Geel, Belgium. maria.de-la-calle@cec.eu.int
Bordin
G
G
Rodriguez
A R
AR
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000295:instrumentation* / Q000379:methods*; D005240:Feasibility Studies; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D020033:Protein Isoforms / Q000097:blood; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D013696:Temperature; D014168:Transferrin / Q000737:chemistry / Q000302:isolation & purification*
D020033:Protein Isoforms; D014168:Transferrin