title: Quantitative evaluation of lectin-reactive glycoforms of α(1)-acid glycoprotein using lectin affinity capillary electrophoresis with fluorescence detection.
abstract: α(1)-Acid glycoprotein (AGP) was previously shown to be a marker candidate of disease progression and prognosis of patients with malignancies by analysis of its glycoforms via lectins. Herein, affinity capillary electrophoresis of fluorescein-labeled AGP using lectins with the aid of laser-induced fluorescence detection was developed for quantitative evaluation of the fractional ratios of concanavalin A-reactive or Aleuria aurantia lectin-reactive AGP. Labeled AGP was applied at the anodic end of a fused-silica capillary (50 μm id, 360 μm od, 27 cm long) coated with linear polyacryloyl-β-alanyl-β-alanine, and electrophoresis was carried out for about 10 min in 60 mM 3-morpholinopropane-1-sulfonic acid-NaOH buffer (pH 7.35). Addition of the lectins to the anode buffer resulted in the separation of lectin-reactive glycoform peaks from lectin-non-reactive glycoform peaks. Quantification of the peak area of each group revealed that the percent of lectin-reactive AGP is independent of a labeling ratio ranging from 0.4 to 1.5 mol fluorescein/mol AGP, i.e. the standard deviation of 0.5% for an average of 59.9% (n=3). In combination with a facile procedure for micro-purification of AGP from serum, the present procedure, marking the reactivity of AGP with lectins, should be useful in determining the prognosis for a large number of patients with malignancies.
authors.affiliation: Department of Natural Sciences, School of Medicine, Fukushima Medical University, Fukushima, Japan. shimurak@fmu.ac.jp
authors.forename: Kiyohito
authors.initials: K
authors.lastname: Shimura
authors.forename: Mayumi
authors.initials: M
authors.lastname: Tamura
authors.forename: Tosifusa
authors.initials: T
authors.lastname: Toda
authors.forename: Shin
authors.initials: S
authors.lastname: Yazawa
authors.forename: Ken-ichi
authors.initials: K
authors.lastname: Kasai
mesh_terms: D015415:Biomarkers / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D003208:Concanavalin A / Q000378:metabolism*; D019075:Electrophoresis, Capillary / Q000379:methods*; D006801:Humans; D037102:Lectins / Q000378:metabolism*; D009961:Orosomucoid / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism
chemical_list: D015415:Biomarkers; D037102:Lectins; D009961:Orosomucoid; C064549:lectin, Aleuria aurantia; D003208:Concanavalin A