Class A 𝜷-Lactamases as Versatile Scaffolds to Create Hybrid Enzymes: Applications from Basic Research to Medicine

Figure 3

BHPs as unique tools for epitope mapping. Schematic representation of the epitope mapping procedure developed by Bannister et al. and Chevigné et al. [8, 11]. The steps for which the enzymatic activity of BlaP, as a reporter or as factor for selection, is critical are highlighted in red. BlaP-Cter: C-terminal sequence of BlaP, starting at the insertion site. BlaP-Nter: N-terminal sequence of BlaP ending at the insertion site. Anti-Ag mab: monoclonal antibody specific to the targeted antigen. Following the elution of phages after the last round of panning, a peptide array can be carried out to verify that the entire antigen sequence is well represented in the phage library. Alanine scanning mutagenesis can be carried out to determine which residues of the minimal sequence encompassing the epitope is in direct contact with the antigen. Figure adapted from Chevigné et al., 2007 [11].