Fig. 10.

Evaluation of amounts of intracellular human CRT protein in yeast cells. a SDS–PAGE of crude yeast lysates and indicated amounts of purified CRT; b, c two Western blot experiments using monoclonal antibodies against human CRT. M prestained protein ladder (ThermoScientific, cat. no. 26616). pFDC, pFDC-CRT and PpB-CRT—crude yeast lysates (10 μg of whole cell protein in each lane). pFDC and pFDC-CRT indicate samples from S. cerevisiae cells transformed with pFDC vector and pFDC-CRT plasmid, respectively. PpB-CRT sample was taken from fed-batch fermentation of high-density P. pastoris culture at 111 h induction time in methanol fed-batch stage (143 h fermentation time). 50, 75, 100, 150, 200, 300—amounts in nanograms of purified S. cerevisiae-secreted human CRT protein loaded on gel