Recombinant Poly p 5 Purification

The allergen rPoly p 5 was acquired, as demonstrated by Bazon et al. [1 (link)]. Briefly, the rPoly p 5 cDNA was cloned in the pPICZαA vector and expressed in Komagataella phaffii (Pichia pastoris) X-33 cells. The soluble rPoly p 5 samples were subjected to purification by application in a prepacked column HisTrap HP™ (Ni⁺² Sepharose™ High Performance; GE Healthcare, Danderyd, Sweden), according to the manufacturer’s instructions. Afterwards, SDS-PAGE electrophoresis tests (12%) were performed to assess the purification process effectiveness. The purified protein was resuspended in phosphate-buffered saline (PBS, pH 7.4) for cell culture experiments.