Figure 3. Analysis of the purity of Asp-NAT from HPLC size exclusion rechromatography. Detector tracing of elution profile of Asp-NAT in HPLC size exclusion re-chromatography (solid line, OD 280 nm) and the composition of proteins as detected by native-PAGE (Tris-acetate, 3-8%) and SDS-PAGE (Tris-glycine, 412%). A semi-preparative HPLC column of ~100 ml capacity (Shodex KW 2004) was used and elution was carried out at 1.0 ml/min collecting 2 ml/fraction. The single peak showed a single band in native gel but multiple bands on SDS-PAGE. Asp-NAT activity (broken line with symbols) corresponded to the peak of OD tracing.
Was this article helpful?