FIGURE 4.

Purity analysis of TrMgTx produced by the two-step purification protocol. (A) Western blot of the sample eluted from the RP-HPLC column (Figures 3C,D) using HRP conjugated anti-His primary antibodies. Lane M: low-molecular-weight protein marker, lane 1: TrMgTx after RP-HPLC purification. (B) TrMgTx eluted from the RP-HPLC column (Figures 3C,D) was loaded on C₁₈ analytical column and eluted with a gradient of 10–30% acetonitrile over 25 min (dotted line, right axis). The absorbance was measured at 280 nm (left axis). Single peak indicates TrMgTx. Purity was calculated as [(area under the peak of interest)/(cumulated area under all peaks) × 100] and is shown in Table 1.