Lectin-based glycoproteomic profiling reveals salivary N-glycan changes recognized by Datura stramonium agglutinin in hepatitis B-related cirrhosis.

Diagnosis of liver cirrhosis using salivary biomarkers offers a non-invasive and cost-effective approach. Our previous lectin microarray screening found an upregulation of Datura stramonium agglutinin (DSA)-recognized salivary glycopatterns in cirrhosis patients. Based on this, a comprehensive glycoproteomic study was conducted. In total, 120 saliva samples from 30 healthy volunteers (HV), 30 patients with hepatitis B (HB), 30 with hepatic cirrhosis (HC), and 30 with hepatocellular carcinoma (HCC) were individually constructed into saliva microarray to analyze the expression differences of DSA-recognized glycopatterns. Then, samples were pooled by group and subjected to lectin blotting to validate differences. The specific glycoproteins were pull-down by DSA-magnetic nanoparticle conjugates for LC-MS analysis, while released glycans were identified by MALDI-TOF-MS. Saliva microarray analysis revealed that DSA-recognized glycopatterns were significantly higher in the HC group than in the other three groups (P < 0.05). A total of 56 glycoproteins were identified, with N-/O-glycan peaks identified in each group as follows: 9/8 (HV), 10/12 (HB), 13/10 (HC), and 12/7 (HCC). According to the specificity of DSA, there are five multi-antennary N-glycans which were overexpressed in HC. These findings provide new insights into cirrhosis-associated glycoproteomic alterations and support the potential development of saliva-based diagnostic strategies.