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| Construction of Small Single Domain Antibody Library Based on Computational Biology and Screening of Anti-estradiol Antibody |
| LIU Chao,QU Xiaochen, WANG Chong,NING Baoan*, GAO Zhixian* |
| Institute of Health and Environmental Medicine, Academy of Military Medical Science, Tianjin 300050, China |
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Abstract Objective To provide new method for screening the specific antibody against small molecular pollutant in food by constructing camelid single domain antibody (VHH) physical library and virtual library.Methods The single domain antibody gene library was constructed by gene cloning, sequencing, homology analysis, the 3D structure virtual library was constructed by secondary structure analysis and homology modeling, and the molecular docking analysis was used to screen the anti-estradiol (E2) antibody in silico.Results The constructed small single domain antibody library contained 2000 antibody strains, and the genetic background, protein structure and the information of complementarity determining region (CDR) of every antibody strain was clear. The binding affinity of the best anti-estradiol VHH screened by molecular docking was 9.56. The dedicated domain of VHH interacted with estradiol was found in the loop structure of CDR. The 50% inhibition concentration (IC50) for E2 was 20 ng/ml, and the limit of detection (LOD, IC10) was 4.097 ng/ml tested by indirect ELISA. Conclusion The camelid single domain antibody library was constructed by computational biology and genetic engineering techniques from which the specific VHH against E2 was screened with clear background and relatively high affinity.
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Published: 30 August 2016
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2016, Vol. 34 
