Expression of functional single-chain variable domain fragment antibody (scFv) against mycotoxin zearalenone in Pichia pastoris.
A synthetic gene coding for single-chain variable domain fragment antibody against mycotoxin zearalenone (scFv-ZEN) has been designed, constructed and expressed in Pichia pastoris. The native scFv-ZEN sequence was optimized to Pichia preference codon usage. The expression level of codon-optimized scFv-ZEN was slightly higher than that of native scFv-ZEN, and its maximum yield reached 328 mg total protein/l in flask culture. The binding activities of two selected clones to ZEN using surface plasmon resonance analysis were comparable or better than that of monoclonal antibody. Our results demonstrate the potential of soluble scFv-ZEN for developing a rapid and affordable immunoassay for detection of ZEN in food and feedstuff.
Chang
Hyun-Joo
HJ
Korea Food Research Institute, 516, Backhyun, Bundang-gu, Sungnam, Kyonggi-do 463-746, South Korea.
Choi
Sung-Wook
SW
Chun
Hyang Sook
HS
D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D004274:DNA, Recombinant; D004591:Electrophoresis, Polyacrylamide Gel; D007135:Immunoglobulin Variable Region / Q000276:immunology*; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D020349:Surface Plasmon Resonance; D015025:Zearalenone / Q000235:genetics / Q000276:immunology*
D004274:DNA, Recombinant; D007135:Immunoglobulin Variable Region; D015025:Zearalenone