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Proteins, Expression, Isolation and Analysis | … The soluble and insoluble fractions were separated by centrifugation at 12,000 rpm for 20 min at 4°C. Then the soluble fraction was purified by using the Glutathione Resin GST Fusion Protein Purification Kit according to the manufacturer's instructions (Genscript cat … | Get A Quote |
Epithelial cell adhesion molecule (EpCAM) is a type I transmembrane glycoprotein overexpressed in human epithelioma but with relatively low expression in normal epithelial tissues. To exploit this differential expression pattern for targeted cancer therapy, an EpCAM-targeted immunotoxin was developed and its antitumor activity was investigated in vitro. An immunotoxin (scFv2A9-PE or APE) was constructed by genetically fusing a truncated form (PE38KDEL) of Pseudomonas aeruginosa exotoxin with an anti-EpCAM single-chain variable fragment (scFv). ELISA and flow cytometry were performed to verify immunotoxin (scFv2A9-PE or APE) antigen-binding activity with EpCAM. Cytotoxicity was measured by MTT assay. Confocal ... More