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.
刘超, 曲晓辰, 王冲, 宁保安, 高志贤. 基于计算生物学方法的小容量单域抗体库的构建及食品中雌二醇特异性抗体的筛选[J]. 解放军预防医学杂志, 2016, 34(3): 308-311.
LIU Chao,QU Xiaochen, WANG Chong,NING Baoan, GAO Zhixian. Construction of Small Single Domain Antibody Library Based on Computational Biology and Screening of Anti-estradiol Antibody. Journal of Preventive Medicine of Chinese People's Liberation Army, 2016, 34(3): 308-311.
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