Isolation of single chain variable fragments against six esters of pyrethrins by subtractive phage display.
Biosci Biotechnol Biochem
; 73(7): 1541-9, 2009 Jul.
Article
em En
| MEDLINE
| ID: mdl-19584536
ABSTRACT
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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Bases de dados:
MEDLINE
Assunto principal:
Piretrinas
/
Região Variável de Imunoglobulina
/
Biblioteca de Peptídeos
/
Ésteres
Tipo de estudo:
Prognostic_studies
Limite:
Animals
Idioma:
En
Revista:
Biosci Biotechnol Biochem
Assunto da revista:
BIOQUIMICA
/
BIOTECNOLOGIA
Ano de publicação:
2009
Tipo de documento:
Article
País de afiliação:
China