Selection and Characterization of Human Anti-MAGE-A1 scFv and Immunotoxin

Abstract

Melanoma-associated antigen (MAGE) is expressed on the surface of multiple tumor cell types and is a promising target of biotherapeutic drug delivery via the anti-MAGE-A1 antibody. In this study, a human single-chain variable fragment (scFv) antibody phage library was generated and applied to recombinant MAGE-A1-coated immunotubes by phage display technology. The soluble anti- MAGE-A1 scFv was expressed and purified by immobilized metal-chelated affinity chromatography (IMAC). The anti-MAGE-A1 scFv could bind native MAGE-A1 confirmed by enzyme-linked immunosorbent assay (ELISA), dot blot, and immunoprecipitation (IP) analysis. The immunotoxin was expressed and purified by IMAC successfully. The results indicated that the human anti-MAGE-A1 immunotoxin could provide a valuable drug for clinic cancer therapy.