Fig. 1: Comparison of glycosylation status in Hp between SPs and HCs.

a AAL blotting of the sera from sepsis patients (n = 121) and healthy donors (n = 73). Statistical significance was calculated with an two-sided unpaired t-test and presented as means ± SD. b,c Lectin blotting for Hp purified from human sera with AAL and PHAL. The membrane was reblotting for β-Hp using anti-Hp antibody. Statistical significance was calculated with an two-sided unpaired t-test and presented as means ± SD. d The top panels show base peak chromatogram (BPC) with structures of major N-glycans observed in this analysis using LC(GC)-ESI MS. The middle panels show extracted ion chromatogram (EIC) of mono-fucosylated tri-antennary N-glycan labeled with aTMT (m/z 818.678 ± 6 ppm) of Hp samples (these are representative data for HC-Hp (HC2) and SP-Hp (SP32)). The bottom panels show average mass spectra (Ave. MS) during 46.0-53.0 min in BPC of the top panels. Structures, abbreviations and theoretical mass for N-glycans labeled with aTMT are shown in Supplementary Fig. 1d. e Relative abundances (%) of non-fucosylated and fucosylated N-glycans labeled with aTMT obtained from LC(GC)-ESI MS analyses of Hp samples (n = 3 for HC-Hp and n = 6 for SP-Hp). Statistical significance was calculated with a two-sided unpaired t-test and presented as means ± SD. f, Relative abundances (%) of bi-, tri- and tetra-antennary N-glycans labeled with aTMT after categorizing terminal- and core-fucosylation obtained from LC(GC)-ESI MS analyses of Hp samples (n = 3 for HC-Hp and n = 6 for SP-Hp). Statistical significance was calculated with a two-sided unpaired t-test and presented as means ± SD. g The top panels show BPC of LC(ODS)-ESI MS analysis. The middle two panels show EIC of peptide (amino acid sequence is NATAK for site 3) containing non- and mono-fucosylated tri-antennary N-glycan (m/z 831.336 and 880.021 ± 6 ppm for 3-0 and 3-F1, respectively) of Hp samples (these are representative data for HC-Hp (HC2) and SP-Hp (SP32)). The bottom panels show average mass spectra (Ave. MS) during 2.6–3.3 min in BPC of the top panels. Structures, abbreviations and theoretical mass for desialo-glycopeptides on each site are shown in Supplementary Fig. 2a. h Relative abundances (%) of non-fucosylated and fucosylated N-glycans on glycopeptide at each site obtained from LC(ODS)-ESI MS analyses of Hp samples (n = 3 for HC-Hp and n = 6 for SP-Hp). Statistical significance was calculated with a two-sided unpaired t-test and presented as means ± SD. i, Relative abundances (%) of bi-, tri- and tetra-antennary N-glycans on glycopeptide at each site obtained from LC(ODS)-ESI MS analyses of Hp samples (n = 3 for HC-Hp and n = 6 for SP-Hp). Statistical analysis was performed using the two-tailed Student’s t-test (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001). ns, not significant.