Fig. 5.

Fine mapping of the target epitope for E184L mAb 1A10 by sequential amino acid deletion from the N and C-terminus of the peptide ¹¹⁰RLCKVFSIMIQRQGFL¹²⁵. A) Analysis of the reactivity of the truncated peptides (R3 peptides) to mAb 1A10 using Western blot. The MBP fused peptides were probed with 1A10 mAb. The mAb-1A10 reacts to peptide R3–1 to R3–9, but not to peptide R3–10 to R3- 14 which indicates the core amino acid for these antibodies lay between ¹¹⁹IQRQGFL¹²⁵. B) Dot blot assay to further confirm the linear core amino acid sequence of mAb 1A10. After succseful expresion of E184L-R3 peptides (R3–1 to R3–12), 2 μL form each was spoted into nitrocellulose membrane and tested against mAb 1A10. The full length E184L and MBP pprotein were used as postive and negative control and are indicated as “PC” and “NC”, respectively. C) Schematic design showing the sequential deletion of amino acid and its respective result of Western blot after staining with mAb 1A10. MBP antibody was used to test the successful expression of each peptide.