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Development of a fast and simple immunochromatographic method to purify alpha 1-acid glycoprotein from serum for analysis of its isoforms by capillary electrophoresis

AutorOngay, Sara ; Lacunza, Izaskun; Diez-Masa, Jose Carlos; Sanz Perucha, Jesús ; Frutos, Mercedes de
Palabras claveSerum purification
Albumin depletion
Biomarker
Glycoprotein
Isoforms
AGP isolation
Fecha de publicación24-mar-2010
EditorElsevier
CitaciónAnalytica Chimica Acta 663 (2) : 206-12 (2010)
ResumenAlpha 1-acid glycoprotein (AGP) is a very heterogeneous glycoprotein presenting several isoforms due to variations in its polypeptidic and glycosidic moieties. Differences in AGP isoforms between healthy and diseased individuals have been related to different pathological situations such as cancer or car- diovascular diseases, among others. Capillary electrophoresis study of the role of AGP isoforms as biomarkers requires prior purification of AGP from biological samples. Current AGP purification methods are time-and labour-consuming, and generally they have not been proven to be compatible with capil- lary electrophoresis analysis. In this work, different methods for AGP purification from human serum are developed and compared. The applicability of acidic precipitation and immunoaffinity chromatographic methods for AGP purification are studied. Two different immunoaffinity approaches are employed; in the first one, interferents present in the AGP sample are captured and removed, and in the second one, AGP is retained in a house-made anti-AGP column, being in this way isolated from the rest of interfer- ents of the sample. Best results in AGP purification from human serum to be analyzed by capillary zone electrophoresis (CZE) were obtained when acidic purification was combined with immunoaffinity chro- matography (IAC) employing the house-made anti-AGP column. The method was shown not to alter the proportion of AGP peaks due to isoforms existing in AGP samples. The applicability of this fast and easy purification method developed for analyzing by CZE isoforms of AGP from natural serum samples by CZE is demonstrated.
Versión del editorhttp://dx.doi.org/10.1016/j.aca.2010.01.054
URIhttp://hdl.handle.net/10261/43158
DOI10.1016/j.aca.2010.01.054
ISSN0003-2670
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