Isolation of single chain variable fragments against six esters of pyrethrins by subtractive phage display.
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.
Fu
Yong-Yao
YY
Genetic Engineering Research Center, Bio-Engineering College, Chongqing University, Chongqing, China.
Li
Zheng-Guo
ZG
Yang
Ying-Wu
YW
Deng
Wei
W
Duan
Wei
W
Miao
Qing
Q
Wang
Zhong-Kang
ZK
Xia
Yu-Xian
YX
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D004952:Esters / Q000737:chemistry*; D007106:Immune Sera / Q000276:immunology; D007114:Immunization; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry; D007147:Immunoglobulin Light Chains / Q000737:chemistry; D007135:Immunoglobulin Variable Region / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D051379:Mice; D008969:Molecular Sequence Data; D019151:Peptide Library; D016133:Polymerase Chain Reaction; D011722:Pyrethrins / Q000737:chemistry* / Q000276:immunology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D017422:Sequence Analysis, DNA
D004952:Esters; D007106:Immune Sera; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D011722:Pyrethrins; D011994:Recombinant Proteins