Figure 5. Epitope mapping of CRP specific scFv LA13-IIC3, LA13-IID4, LA13-IIE3. A series of overlapping 15mer oligopeptides covering the whole CRP protein sequence with an offset of three amino acids was synthesized onto filter membranes. These filter membranes were subsequently immunostained with CRP specific scFv antibodies LA13-IIC3 (A), LA13-IID4 (B), and LA13-IIE3 (C). Bound scFv antibodies were detected with α-myc-tag antibody and secondary antibody AP conjugate. Both epitopes are highlighted (yellow, space fill) in one subunit of the CRP pentamer structure (D). Epitope location based on structure (PDB ID: 1B09)⁴: peptide backbone (gray ribbon), side chains (lines), and polyhendron surface (points) are shown (visualization with 3D-Mol Viewer), epitopes indicated in yellow. Important amino acid residues of the epitopes identified by an additional alanine walk (data not shown) are underlined (* only IIC3)