Figure 1

SDS-PAGE analysis of the product obtained during the different steps during the purification of the IgE hyporeactive (K8A, N11A, D55A) (A) and wild-type allergen (B) C-terminal domain of Phl p 1.0102. The following fractions were analysed: Lysate (L), soluble fraction (S), column flow through (FT) and wash (W) fractions, eluted fractions (pool), the pool after digestion with PreScission protease (+3C), and the final, purified protein (final). Dilutions of the samples prior to electrophoresis are indicated above the lanes. The purified non-GST fused IgE hyporeactive domain, depicted to the right, migrates as a single band, although at a slightly higher apparent molecular weight (14 kDa) than expected (11 kDa). The purified GST-coupled protein (37 kDa) as well as GST (26 kDa) are clearly visible with expected molecular weights.