Wheat germ cell-free system-based production of hemagglutinin-neuraminidase protein of human parainfluenza virus type 3: generation and characterization of monoclonal antibody
| Autor: | Ayumi Kudoh, Satoko Matsunaga, Hirokazu Kimura, Hisashi Hirano, Akiko Okayama, Izumi Matsuo, Hideaki Shimizu, Shiho Kawakami, Akihide Ryo, Naoki Yamamoto, Yuki Matsushima, Nobuhiko Okabe, Hiroyuki Tsukagoshi |
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| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Microbiology (medical)
Proteomics Monoclonal antibody Immunoprecipitation medicine.drug_class lcsh:QR1-502 Human parainfluenza virus 3 Microbiology Virus lcsh:Microbiology law.invention law medicine Original Research Article HN Protein Cell fusion biology Virology Molecular biology cell-free protein synthesis biology.protein Recombinant DNA hemagglutinin-neuraminidase protein Antibody Clone (B-cell biology) |
| Zdroj: | Frontiers in Microbiology, Vol 5 (2014) Frontiers in Microbiology |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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