Identification of antibody isotypes, epitopes and cross reactivity to mammalian recombinant PrP.

Antibodies which were shown to have similar binding motifs were assigned to distinct groups (1 to 4, whereby the common amino acid sequences are highlighted in bold/underlined text). For comparison, the epitopes and source of commercially available PrP-antibodies (Sha 31, P4 and 6H4) are indicated. The amino acid sequence of the core binding region of each antibody is indicted. The epitopes for ROS-JC4, ROS-EA6 and ROS-FD12 could not be determined (ND) using Pepscan, whereas ROS-EC9 and ROS-IH11 were not tested (NT) in pepscan analysis. Several mammalian species of recombinant PrP (typically 25 ng protein) were resolved using SDS-PAGE and probed with the mAbs post Western transfer. ‘+’ indicated antibodies which cross react and ‘–’ antibodies that did not cross react. ROS-HC7, Sha 31, P4, 6H4 and 12B2 were not tested (NT) in this assay.