| 19584536 |
Fu YY, Li ZG, Yang YW, Deng W, Duan W, Miao Q, Wang ZK, Xia YX: Isolation of single chain variable fragments against six esters of pyrethrins by subtractive phage display. Biosci Biotechnol Biochem. 2009 Jul;73(7):1541-9. Epub 2009 Jul 7.
Recombinant antibodies (rAbs) are a new diagnostic test for immulogical detection. To date, there are no reports about anti-pyrethrins rAbs. Here we describe the generation of monomeric and dimeric single chain variable fragments (scFvs) with affinity for six esters of pyrethrins using a subtractive phage display technology. First, scFv libraries with long-linker (Ger (4) Ser)(3) and short-linker (Ger (4) Ser) were established to contain 1.04 x 10 (7) or 6.07 x 10 (6) transformants. After four rounds of panning, phage ELISA demonstrated that three clones (E2, F2, and H7) showed higher affinity from the long-linker library, and clones (h6, a5) exhibited better antibody activity to pyrethrin I and II from the short-linker library. The scFv candidates were sequenced to identify the specific antibody response against pyrethrins. Isolated scFvs constitute valuable tools for real-time detection of pyrethrins. In addition, the subtractive phage display provides a simple approach for isolation of scFvs. |
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