Comparative Analysis for Glycopatterns and Complex-Type N-Glycans of Glycoprotein in Sera from Chronic Hepatitis B- and C-Infected Patients

Abstract

Background: Chronic infection with HBV (CHB) or HCV (CHC) is the most common chronic viral hepatitis that can lead to cirrhosis and hepatocellular carcinoma in humans, their infections have distinct pathogenic processes, however, little is known about the difference of glycoprotein glycopatterns in serum between hepatitis B virus (HBV)- and hepatitis C virus (HCV)-infected patients.Methods: A method combining the lectin microarrays, letin-mediated affinity capture glycoproteins, and MALDI-TOF/TOF-MS was employed to analyze serum protein glycopatterns and identify the glycan structures from patients with CHB (n = 54) or CHC(n = 47), and healthy volunteers (HV, n = 35). Lectin blotting was further utilized to validate and assess the expression levels of their serum glycopatterns. Finally, the differences of the glycoprotein glycopatterns were systematically compared between CHB and CHC patients.Conclusions: As a result, there were 11 lectins (e.g., HHL, GSL-II, and EEL) exhibited significantly increased expression levels, and three lectins (LCA, VVA, and ACA) exhibited significantly decreased expression levels of serum protein glycopatterns only in the CHB patients. However, DBA exhibited significantly decreased expression levels, and two lectins (WGA and SNA) exhibited significantly increased expression levels of serum glycopatterns only in the CHC patients. Furthermore, LEL and MAL-I showed a coincidentally increasing trend in both CHC and CHB patients compared with the HV. The individual analysis demonstrated that eight lectins (MPL, GSL-I, PTL-II, UEA-I, WGA, LEL, VVA, and MAL-I) exhibited a high degree of consistency with the pooled serum samples of HV, CHB, and CHC patients. Besides, a complex-type N-glycans binder PHA-E+L exhibited significantly decreased NFIs in the CHB compared with HV and CHC subjects (p < 0.01). The MALDI-TOF/TOF-MS results of N-linked glycans from the serum glycoproteins isolated by PHA-E+L-magnetic particle conjugates showed that there was an overlap of 23 N-glycan peaks (e.g., m/z 1419.743, 1663.734, and 1743.581) between CHB, and CHC patients, 5 glycan peaks (e.g., m/z 1850.878, 1866.661, and 2037.750) were presented in virus-infected hepatitis patients compared with HV, 3 glycan peaks (1460.659, 2069.740, and 2174.772) were observed only in CHC patients. Our data provide useful information to find new biomarkers for distinguishing CHB and CHC patients based on the precision alteration of their serum glycopatterns.

Integrated Glycome Strategy for Characterization of Aberrant LacNAc Contained N-Glycans Associated With Gastric Carcinoma

“…The N-glycans were released by PNGase F glycosidase (New England Biolabs, Beverly, MA) according to previous protocols (25, 26). Briefly, 200 μg of isolated glycoproteins were concentrated and desalted by adding to a size-exclusion spin ultrafiltration (Amicon Ultra-0.5 mL 10,000 MW cut off, Millipore).…”

“…The MAL-I-enriched N-linked glycans were characterized using MALDI-TOF/TOF-MS (UltrafleXtreme, Bruker Daltonics, Germany) as previously described (25, 26). Briefly, the glycans were resuspended in 10 μL of methanol, then glycan samples and DHB were spotted on a MTP AnchorChip, respectively to recrystallize the glycans.…”

Roles of glycoprotein glycosylation in the pathogenesis of an endemic osteoarthritis, Kashin–Beck disease, and effectiveness evaluation of sodium hyaluronate treatment

“…The mixture was then used in the lectin microarrays. Finally, incubation was performed in the chamber at 37 °C for 3 h [9].…”

“…The membrane was incubated with Cy5-labeled (GE Healthcare, Buckinghamshire, UK) lectins (2 mg/) at 4 °C overnight in the dark. Finally, the membrane was scanned using a phosphorimager (Molecular Dynamics Inc., Sunnyvale, CA, USA) [9].…”

Clinical application of quantitative glycomics

“…N-glycans profiles by MALDI-TOF/TOF-MS have shown that glycans F1N5H4 (bisecting), N5H5 (bisecting) and F1N6H3F1 are overexpressed in CHB patients. In contrast, glycans F1N5H4 (bisecting), N3H5, N6H5 and F1N5H6 are overexpressed in CHC patients, compared to healthy individuals [194]. The over-expression of bisecting glycan F1N5H4 was also observed in liver cancer.…”