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《Medical Journal of Chinese People's Liberation Army》 2010-01
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Screening of mimotopes of swine influenza virus A (H1N1) by phage display technology

CAI Jiong,ZHONG Yan-wei,JING Xia,CHEN Li,LI Fang,XU Dong-ping. Department of Nuclear Medicine,PUMC Hospital,PUMC & CAMS,Beijing 100730,China  
Objective To screen the specific antigen mimotopes of influenza A(H1N1)virus by phage display technology,in order to pave a way to develop novel influenza vaccine.Methods Using human convalescent serum of pandemic influenza A(H1N1)in 2009 as solid-phase selective molecule,an artificial synthesized phage randomly displaying cyclic 7-mer peptide was screened with three rounds of "absorption-elution-amplification" selection.At the end of the third round selection,32 clones were randomly chosen from the top-agar phage plaques and placed onto E.coli ER2378 in logarithmic growth phase.After culturing for 5 hours,the positive clones were identified by enzyme linked immunosorbent assay(ELISA),cross reaction test and competitive inhibition assay.The identified positive clones were analyzed by DNA sequencing.The decoded amino acids sequences,which displayed on the surface of phage,were aligned with the hemagglutinin(HA)gene of influenza virus by homology comparison for definition of the mimotopes of influenza A(H1N1)virus.Results After enriching the specific antibody-binding phages from phage displaying peptide library,12 positive clones were chosen from 32 randomly selected clones.DNA sequencing and homology comparison showed that one epitope PLHARLP was confirmed as mimotope of swine influenza A(H1N1)virus antigen,which was composed of the 52nd,53rd,59th,60th,61st,83rd and 271st amino acid.Conclusions Swine influenza A(H1N1)mimotope has been obtained by cyclic 7-mer peptide phage library screening.This result provides a basis for developing new influenza virus vaccine from virus antigen mimotopes.
【Fund】: 国家自然科学基金(30771993)
【CateGory Index】: R392
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