Abstract
A phage display library of single chain variable fragment (scFv) antibodies was constructed to screen specific scFv antibodies against amatoxins. One recombinant scFv antibody with high specificity for amatoxins, termed scFv-A4, was isolated from the library by biopanning. SDS-PAGE analysis showed that the soluble scFv-A4 protein was expressed successfully, and the protein posed relatively good specificity with an IC50 of 77.48 ng/mL and IC15 of 1.91 ng/mL using indirect competitive ELISA (ic-ELISA). On the basis of the expressed scFv-A4 protein, a rapid and simple new immunoassay using surface plasmon resonance (SPR) sensor for the detection of amatoxins was developed. The IC50 and IC15 of new immunoassay were 7.66 and 0.17 ng/mL. The sensitivity of immunoassay using SPR sensor was about 10 times that of ic-ELISA. These results showed the potential usefulness and advantages of new immunoassay using SPR sensor for the detection of toxins.
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