LC–MS/MS Quantitation
of Esophagus Disease
Blood Serum Glycoproteins by Enrichment with Hydrazide Chemistry and
Lectin Affinity Chromatography
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Abstract
Changes
in glycosylation have been shown to have a profound correlation
with development/malignancy in many cancer types. Currently, two major
enrichment techniques have been widely applied in glycoproteomics,
namely, lectin affinity chromatography (LAC)-based and hydrazide chemistry
(HC)-based enrichments. Here we report the LC–MS/MS quantitative
analyses of human blood serum glycoproteins and glycopeptides associated
with esophageal diseases by LAC- and HC-based enrichment. The separate
and complementary qualitative and quantitative data analyses of protein
glycosylation were performed using both enrichment techniques. Chemometric
and statistical evaluations, PCA plots, or ANOVA test, respectively,
were employed to determine and confirm candidate cancer-associated
glycoprotein/glycopeptide biomarkers. Out of 139, 59 common glycoproteins
(42% overlap) were observed in both enrichment techniques. This overlap
is very similar to previously published studies. The quantitation
and evaluation of significantly changed glycoproteins/glycopeptides
are complementary between LAC and HC enrichments. LC–ESI–MS/MS
analyses indicated that 7 glycoproteins enriched by LAC and 11 glycoproteins
enriched by HC showed significantly different abundances between disease-free
and disease cohorts. Multiple reaction monitoring quantitation resulted
in 13 glycopeptides by LAC enrichment and 10 glycosylation sites by
HC enrichment to be statistically different among disease cohorts