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

[ Vol. 13 , Issue. 8 ]

Author(s):

Hong Lin, Yuan Mao, Da-Wei Zhang, Hu Li, Jin-Rong Qiu, Jin Zhu and Ren-Jie Chen   Pages 1259 - 1266 ( 8 )

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.

Keywords:

Phage display antibody library, anti-MAGE-A1 scFv fragment, immunotoxin.

Affiliation:

, , , , , , Department of Otolaryngology-Head and Neck Surgery, the Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, China.



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