Characterization of an Isotype-Dependent Monoclonal Antibody against Linear Neutralizing Epitope Effective for Prophylaxis of Enterovirus 71 Infection
Figure 3
Detailed epitope mapping of mAb 51 and 53 by Western blot analysis.
(A) C-terminal truncated fragments of VP1 protein (a–f), N-terminal truncated fragments of VP1 protein (aa–hh), and GST-(epitope) peptide spanning regions as depicted. Expression of protein fragments was determined with mAb against GST protein. (B) Western blot results of C-terminal truncated proteins (a–f) indicated interaction of both mAb 51 and 53 with fragment f only, implying that the last amino acid of the epitope should be at amino acid 219 or 220. (C) Western blot results of N-terminal truncated proteins (aa–hh) indicated that the first amino acid of the epitope should be amino acid 214. Thus, the putative epitope of mAb 51 and 53 should span amino acid 214 to either 219 or 220. (D) Western blot analysis of GST-putative epitope proteins (GST-KQEK, GST-HKQEKD, GST-HKQEK, GST-KQEKD) and GST as negative control. Interaction with both GST-HKQEKD and GST-KQEKD indicated that the epitope of mAb 51 and 53 is KQEKD (215–219).