Epitope mapping of anti-AapBrpt1.5 MAbs.

<p>AapBrpt1.5 N-terminally fused with a GB1-tagged six-histidine (GB1-His) tag was truncated into a series of fragments as shown in the schematic diagrams, and the binding ability between the truncated fragments and MAbs was analyzed using immunoprecipitation. (A) Preliminary epitope mapping. (B) Precise recognition site mapping of MAb_25C11 and MAb_20B9. (C) Precise recognition site mapping of MAb_18B6. (D) Domain structures of Aap from <i>S. epidermidis</i> RP62A (“RP62A”) and ATCC 12228 (“12228”). The A-repeat region, the putative globular domain (α/β), the B-repeat region containing 6 or 12 tandem Brpt constructs, the collagen-like proline/glycine-rich region, the domain boundary of AapBrpt1.5, and the MAb epitopes are illustrated. (E) Amino acid sequence alignment of AapBrpt constructs. The AapBrpt construct in AapBrpt1.5 (GenBank NP_763730) and twelve distinct AapBrpt constructs from <i>S. epidermidis</i> RP62A (RP62A 01-12, GenBank YP_189945) were aligned using the ClustalW2 program (<a href="http://www.ebi.ac.uk/Tools/clustalw2" target="_blank">http://www.ebi.ac.uk/Tools/clustalw2</a>). The identified epitopes of the MAbs are shown in boxes, and the identical residues are marked with asterisks. The conserved substitutions are represented by “:”, and semi-conserved substitutions are represented by “.”. Two conserved His residues in AapBrpt constructs are marked with triangles.