FIG. 1.

EpCAM16 was successfully purified from bacterial inclusion bodies. (A) A diagram shows the plasmid p16xEpCAM.pET21 containing the EpCAM16 construct consisting of an Nco1 restriction site, the VH and VL regions of anti-human CD16 (NM3E2), a segment of human muscle aldolase, the VH and VL regions of humanized anti-EpCAM (MOC31), and a XhoI restriction site. (B) The absorbance trace of the refolded protein eluted from the Q-sepharose ion-exchange column using a stepwise NaCl gradient. (C) The gel stained with Coomasie blue showing a single band of about 54 kDa eluted from the second column that we ran (size-exclusion column). Lane 1, molecular-weight standards; Lane 2, nonreduced EpCAM16; Lane 3, nonreduced anti-Ly5.1 monoclonal antibody control. EpCAM, epithelial cell adhesion molecule.