Applied Biochemistry and Biotechnology

, Volume 169, Issue 2, pp 502–510 | Cite as

Il17A Autoantibody Induced by Recombinant Protein Ag85A-Il17A

  • Rong Jin
  • Sheng Guo
  • Liangxia Wu
  • Xiaoyong Fan
  • Hui Ma
  • Douglas B. Lowrie
  • Jianhua Zhang
Article

Abstract

IL17A is a widely studied cytokine which plays an important role in autoimmune diseases as well as asthma. In this study, murine IL17a gene fragment was inserted into pET28a-Ag85A and it was expressed as a fusion protein named Ag85A-IL17A. Recombinant protein Ag85A-IL17A was expressed as an open reading frame of 1,287 bp, encoding a polypeptide of 429 amino acid residues with a predicted molecular mass of 50 kDa. It was expressed in Escherichia coli. BL21 and purified by metal affinity chromatography using a nickel–nitrilotriacetic acid column. Bioinformatics revealed that Ag85A-IL17A contained B cell epitopes of IL17A. Purified proteins Ag85A-IL17A and Ag85A were injected with adjuvants into mice then antibody responses in sera were measured by enzyme-linked immunosorbent assay and western blot assay. Experiments showed that purified protein Ag85-IL17A could bind to a standard IL17A antibody and could induce IL17A autoantibody in mice.

Keywords

IL17A Ag85A Cloning Expression Autoantibody B cell epitope 

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Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  • Rong Jin
    • 1
  • Sheng Guo
    • 1
  • Liangxia Wu
    • 1
  • Xiaoyong Fan
    • 1
  • Hui Ma
    • 1
  • Douglas B. Lowrie
    • 1
  • Jianhua Zhang
    • 1
  1. 1.ShanghaiChina

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