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. Author manuscript; available in PMC: 2015 Feb 13.
Published in final edited form as: Biochem Biophys Res Commun. 2013 Jan 9;431(2):215–220. doi: 10.1016/j.bbrc.2012.12.132

Figure 1. Analysis of the immunogenicity of the glycoforms of AniA.

Figure 1

(A) Structure of the N. meninigitidis O-linked trisaccharide molecule Gal (β1–4) Gal (α1–3) 2,4-diacetamido-2,4,6-trideoxyhexose (DATDH). The arrows indicate the truncations of the trisaccharide structure in pglA and pglL mutant strains. (B) A typical result from western blot analyses of antisera from mice immunised with AniA glycosylated with the DATDH monosaccharide against outer membrane proteins from C311 (AniA glycosylated with a trisaccharide molecule), C311 pglA (AniA glycosylated with a monosaccharide) and C311 pglL (non-glycosylated AniA). AniA is detected at approximately 51kDa in C311 and at slightly lower molecular weights in C311 pglA and in C311 pglL due to glycosylation with the monosaccharide and loss of the glycan, respectively. (C) A typical result from western blot analyses of antisera from mice immunised with non-glycosylated AniA against outer membrane proteins from C311, C311 pglA and C311 pglL. (D) A typical result from western blot analyses of antisera from mice immunised with truncated AniA lacking the C-terminal glycosylation region (1: 100) against outer membrane proteins from C311, C311 pglA and C311 pglL. (E) Western blot analysis of outer membrane proteins from C311, C311 pglA and C311 pglL using an anti-AniA mAb to show that equal amounts of the AniA protein were present in each sample. (F). Immunisation with AniA lacking the C-terminus elicits a greater antibody response against the non-glycan epitopes of the protein. The reciprocal of the antibody titres from mice immunised with AniA glycosylated with the monosaccharide (AniA pglA-), non-glycosylated AniA (AniA pglL-) and AniA lacking the C-terminus (Truncated AniA) against recombinant truncated AniA (Antigen 5) are shown. Error bars represent ± 1 standard deviation from the mean. A statistically significant difference was detected between the mean antibody titre of the group immunized with AniA pglA- and the group immunised with Truncated AniA (p = 0.0433).