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Construction of scFv phage display library with hapten-specific repertories and characterization of anti-ivermectin fragment isolated from the library

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Abstract

A specific phage display library was developed for screening antibodies against micro-molecular substances with 16-membered macrocyclic backbone. Through mRNA extraction, RT-PCR and gene splicing by overlap extension PCR (SOE-PCR), single-chain fragment variable (scFv) gene fragments about 750 bp were generated and ligated with the phagemid pCANTAB5E and were transformed into competent cells. A phage display library containing 2.4 × 106 clones was constructed from milbemycin oxime-bovine serum albumin (MILO-BSA) immunized mice. The screening was carried out by ivermectin-bovine serum albumin (IVM-BSA) with different concentration levels. Furthermore, scFv phage clones were isolated within the four round library panning and screening by phage- ELISA, 10 positive clones were obtained finally. These positive clones were then sequenced, and their soluble type antibodies were identified and showed significant binding activity.

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Acknowledgments

The authors are grateful for support from the Chinese Academy of Sciences special support project (3047115), and 948 Projects of the Ministry of Agriculture (2005-Z24).

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Authors and Affiliations

  1. Institute of Food Quality Safety and Detection Research, Jiangsu Academy of Agricultural Sciences, 210014, Nanjing, China

    Xiao Zhang, Cunzheng Zhang, Yuan Liu, Xiangyang Yu & Xianjin Liu

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  1. Xiao Zhang
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  2. Cunzheng Zhang
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  3. Yuan Liu
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  4. Xiangyang Yu
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  5. Xianjin Liu
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Correspondence to Xianjin Liu.

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Zhang, X., Zhang, C., Liu, Y. et al. Construction of scFv phage display library with hapten-specific repertories and characterization of anti-ivermectin fragment isolated from the library. Eur Food Res Technol 231, 423–430 (2010). https://doi.org/10.1007/s00217-010-1293-8

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  • DOI: https://doi.org/10.1007/s00217-010-1293-8

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