Selection and characterization of human anti-MAGE-A1 scFv and immunotoxin.
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.
Jiangsu Provincial Blood Center, Nanjing 210042, China.
Hong
H
Lin
Yuan
Y
Mao
Da-Wei
DW
Zhang
Hu
H
Li
Jin-Rong
JR
Qiu
Jin
J
Zhu
Ren-Jie
RJ
Chen
D000036:Abrin / Q000235:genetics; D000595:Amino Acid Sequence; D017209:Apoptosis / Q000187:drug effects; D001483:Base Sequence; D045744:Cell Line, Tumor; D006801:Humans; D000922:Immunotoxins / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D058950:Melanoma-Specific Antigens / Q000276:immunology* / Q000378:metabolism; D008969:Molecular Sequence Data; D019151:Peptide Library; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*
D000922:Immunotoxins; C072011:MAGEA1 protein, human; D058950:Melanoma-Specific Antigens; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies; D000036:Abrin