Figure 3

Address determination of the 2E4 epitope. (a) The 3D structure of the entire B2L protein was generated via the SWISS-MODEL server and contains three selected regions: F1 (aa58-112 indicated in red), F2 (aa 205-23 indicated in blue) and F3 (aa338- 375 indicated in green). (b) Three peptides of interest in B2L. (c) The back view of (b). (d) The separate states of F1, F2 and F3. (e) SDS-PAGE and western blot of the three fragments. The GST-fused peptides F1, F2 and F3 were separated by 12% SDS-PAGE. Western blot analysis with 2E4 corresponded with the fusion proteins shown by PAGE, but none of them showed an obvious band on the nitrocellulose membrane. (f) F1 was found to specifically recognize mAb 2E4 by ELISA. OD450 nm values less than 0.5 were considered non-specific binding to 2E4 except the full-length B2L antigen and F1 in this ELISA assay. (**p < 0.01, represents a statistically highly significant difference; Mean ± SD, n = 3).