Frontiers in Microbiology (May 2014)

Wheat germ cell-free system-based production of hemagglutinin-neuraminidase protein of human parainfluenza virus type 3: generation and characterization of monoclonal antibody

  • Satoko eMatsunaga,
  • Shiho eKawakami,
  • Akiko eOkayama,
  • Hiroyuki eTsukagoshi,
  • Ayumi eKudoh,
  • Izumi eMatsuo,
  • Yuki eMatsushima,
  • Hideaki eShimizu,
  • Nobuhiko eOkabe,
  • Hisashi eHirano,
  • Naoki eYamamoto,
  • Hirokazu eKimura,
  • Akihide eRyo

DOI
https://doi.org/10.3389/fmicb.2014.00208
Journal volume & issue
Vol. 5

Abstract

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Human parainfluenza virus 3 (HPIV3) commonly causes respiratory disorders in infants and young children. Monoclonal antibodies (MAbs) have been produced to several components of HPIV3 and commercially available. However, the utility of these antibodies for several immunological and proteomic assays for understanding the nature of HPIV3 infection remain to be characterized. Herein, we report the development and characterization of monoclonal antibodies against hemagglutinin-neuraminidase (HN) of HPIV3. A recombinant full-length HPIV3-HN was successfully synthesized using the wheat-germ cell-free protein production system. After immunization and cell fusion, 36 mouse hybridomas producing MAbs to HPIV3-HN were established. The MAbs obtained were fully characterized using ELISA, immunoblotting and immunofluorescent analyses. Of the MAbs tested, single clone was found to be applicable in both flow cytometry and immunoprecipitation procedures. By utilizing the antibody, we newly identified HPIV3-HN binding host proteins via immunoprecipitation-based mass spectrometry analysis. This study provides the availability of our newly-developed MAbs as a valuable tool for the study of HPIV3 infection as well as the several diagnostic tests of this virus.

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