Figure 5.

Chromatographic purification of DmAMP1-CRPs and RsAFP2-CRPs from extracellular fluid fraction of Arabidopsis plants transformed with the double-protein construct pFAJ3105. The extracellular fluid fraction was loaded on a C8 RP-HPLC column equilibrated in 15% (v/v) acetonitrile in 0.1% (v/v) trifluoroacetic acid (TFA). The column was eluted at a flow rate of 1 mL min⁻¹ for 15 min with 15% (v/v) acetonitrile in 0.1% (v/v) TFA followed by a linear gradient of acetonitrile in 0.1% (v/v) TFA from 15% (v/v) to 50% (v/v) acetonitrile in 35 min. The eluate was monitored by on-line measurement of the A₂₈₀ (A₂₈₀;—) and the acetonitrile gradient (---) was monitored with an on-line conductivity sensor. Fractions were collected and assessed for the presence of DmAMP1-CRPs and RsAFP2-CRPs by the respective ELISA assays. Dotted bars represent the presence of DmAMP1-CRPs (p3105EF1 and p3105EF2), whereas the black bar represents the presence of RsAFP2-CRPs (p3105EF3). Triangles indicate the elution position of native DmAMP1 (white triangle) and of native RsAFP2 (black triangle).