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《Progress in Veterinary Medicine》 2018-05
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Construction and Screening of Nanobody Library Against BVDV

DING Jin-hua;LI Qi-feng;MA Xu-sheng;XIAO Hong-ran;XIAO Sheng-zhong;WU Peng;SHENG Jin-liang;CHEN Chuang-fu;College of Animal Science and Technology,Shihezi University;Tian Kang Biological Co.Ltd.;College of Life Science,Shihezi University;  
The aim was to obtain the specific nanobodies against bovine viral diarrhea virus(BVDV).Using BVDV-E0 recombinant protein to immunize the alpaca,the lymphocytes in the blood were separated.The phage display library was constructed by using phage display technology,and then the phages combined with BVDV-E0 protein were obtained through three successive bioscai sieves,the obtained VHH sequences were sequenced and alignmented.The high affinity nanobody against BVDV-E0 was screened by ELISA,and then the affinity and activity of the nannobody were verified.The results showed that phage expression library with a insertion rate 86%,the database 1.3×1011 cfu was successfully constructed.Five BVDV-E0 positive clones were obtained by screening,these genes were cloned into prokaryotic expression system,and the high purity BVDV-E0 nanobodies were obtained.Two different high affinity VHH genes were obtained by affinity identification.The high purity BVDV-E0 nanobodies could be combined with artificial antigen E0,and also be blocked by competitive antibodies.Thies study laid a foundation for further study on assembly of BVDV kit and development of BVDV vaccine.
【Fund】: 国家自然科学基金项目(U1303283)
【CateGory Index】: S852.653
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