Production of monoclonal antibodies reactive with a denatured form of the friend murine leukemia virus gp70 envelope protein: Use in a focal infectivity assay, immunohistochemical studies, electron microscopy and western blotting

Michael N. Robertson, Masaaki Miyazawa, Shiro Mori, Byron Caughey, Leonard H. Evans, Stanley F. Hayes, Bruce Chesebro

Research output: Contribution to journalArticlepeer-review

96 Citations (Scopus)

Abstract

Four monoclonal antibodies were selected for their ability to recognize the envelope protein of Friend murine leukemia virus (F-MuLV) in methanol-fixed tissue culture cells. Each of these monoclonal antibodies was found to react only with F-MuLV. By using recombinant retroviruses, it was determined that each of the monoclonal antibodies recognized the C-terminal one-third of the F-MuLV gp70 envelope protein. The monoclonal antibodies were effective in radioimmunoprecipitation of F-MuLV proteins, and one of the antibodies, 720, was also effective in Western blotting. The ability of antibody 720 to react with F-MuLV in methanol-fixed cells facilitated the use of a sensitive immunoperoxidase method with a focal virus infectivity assay. In immunohistochemical studies using light microscopy, antibody 720 could specifically label F-MuLV-infected cells in acetone-fixed tissue sections from F-MuLV-infected animals. Finally, in immuno-gold labelling studies using electron microscopy, antibody 720 could be used to distinguish F-MuLV from amphotropic MuLV.

Original languageEnglish
Pages (from-to)255-271
Number of pages17
JournalJournal of Virological Methods
Volume34
Issue number3
DOIs
Publication statusPublished - 1991 Oct
Externally publishedYes

Keywords

  • Electron microscopy
  • Immunohistochemistry
  • Monoclonal antibody
  • Retrovirus

ASJC Scopus subject areas

  • Virology

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