Prokaryotic expression under different condition and stability of antibody single-chain variable fragment(scFv) against fumonisin B1,a pernicious mycotoxic,were investigated for promoting its application in immunoassay.After construction of expression vector pET22b-1D11 and pMAL-1D11,we analyzed the influence of temperature,concentration of inducer,host bacteria,and high soluble protein tag on scFv expression form.And the thermal stability and organic solvent resistance of scFv were analyzed by enzyme-linked immunosorbent assay(ELISA).The results showed that scFv and its fusion protein were mainly in the formation of inclusion body after optimization of the above conditions.Heat treatment could reduce or even inactivate the scFv activity,and methanol could reduce the sensitivity of scFv-based ELISA.