Liquid chromatographic and mass spectrometric analysis of human serum acid alpha‐1‐glycoprotein

Abstract

Human serum acid alpha-1-glycoprotein (AGP, orosomucoid) content of healthy individuals and cancer patients was measured, isolated and purified using a protocol of fast and biocompatible sample preparation, ion exchange and dye-ligand affinity chromatographic methods. In comparison to the healthy individuals significantly higher serum AGP levels were found in a wide spectrum of cancer patients, indicating its diagnostic value in the malignant disease. Oligosaccharide content of AGP samples was separated following PNGase F enzyme digestion and analysed by RP-HPLC and MALDI-TOF mass spectrometry. RP-HPLC and MALDI-TOF mass spectrometric analysis of sugar constituents of AGP specimen originated from selected cancer patients with high serum AGP levels indicated the appearance of anomal distribution of bi-, tri- and tetra-antennary oligosaccharide structures compared to the healthy controls.

Statistical evaluation of CZE‐UV and CZE‐ESI‐MS data of intact α‐1‐acid glycoprotein isoforms for their use as potential biomarkers in bladder cancer

“…This result would indicate that an increase of tetra-antennary fucosylated glycans in bladder cancer patients when compared to healthy volunteers allows partial sample classification. This result corroborates the discussion performed for PCA and ANOVA results of glycan data analysis, and it is according to the results of Vekey's group [37,38]. However, the indexes employed by them to differentiate among the groups of study [37] were not statistically different between our groups of study.…”

“…However, five out of the six variables correspond to tri-and tetra-antennary glycans which could indicate that the fucosylation on these glycans could be more discriminatory than the fucosylation on biantennary glycans. The discriminatory power suggested for fucosylation and high antennary glycans is in accordance with the study from Vekey's group for other types of cancer [38]. A recent study performed by the same group [37] for the same types of cancer shows that fucose and branching indexes depend on the pathophysiology, but that statistical techniques are needed for correct classification.…”

CIEF with hydrodynamic and chemical mobilization for the separation of forms of α‐1‐acid glycoprotein

“…An AGP sample from a pool of sera of patients with ovary cancer (from now on named as AGP from ovary cancer sample) was donated by Professor Kremmer (National Institute of Oncology, Budapest, Hungary), who purified this sample according to his published method [6].…”

“…However, it will be necessary to analyze a large number of samples with this method to establish solid conclusions. AGP from ovary cancer sample, purified following the method described in [6] also seemed to be better resolved and thus easier to integrate for area studies and reproducibility purposes. The several electrophoretical zones have been assigned, as shown in Fig.…”

Determination of sialic acids released from glycoproteins using capillary zone electrophoresis/electrospray ionization mass spectrometry

“…glycoproteins, consists of a wide variety of neutral sugars, hexosamines and sialic acids. Recent observations indicate that the remarkable variability of these glycan chains in branching and length as well as the terminal groups contain characteristic information of various physiological and pathological properties [1]. Generally, sialic acids -compounds derived from the nine-carbon sugar neuraminic acid (5-amino-3,5-dideoxy-D-glycero-D-galacto-nonulosic acid) -form the terminal end in sugar chains of glycoproteins.…”