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Preparation of rat monoclonal antibodies to epitopes encoded by the viral oncogene (v‐ fms ) of McDonough feline sarcoma virus
Author(s) -
Anderson Soni J.,
Furth Mark E.,
Wolff Linda,
Ruscetti Sandra K.,
Sherr Charles J.
Publication year - 1982
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.240190309
Subject(s) - monoclonal antibody , epitope , biology , virology , microbiology and biotechnology , antibody , immunofluorescence , antigen , virus , oncogene , cell , immunology , biochemistry , cell cycle
The McDonough strain of feline sarcoma virus (SM‐FeSV) contains a viral oncogene, v‐ fms , transduced from cat cellular genetic sequences designated c‐ fms . Monoclonal antibodies reactive to antigenic determinants encoded by v‐ fms were prepared by immunizing rats with live, syngeneic SM‐FeSV‐transformed cells, and fusing splenic lymphocytes from a tumor‐bearing animal with cultured rat myeloma cells. Culture supernatants from hybrids producing antibodies to epitopes encoded by v‐ fms were identified by immunoprecipitation of radiolabeled polypeptides from SM‐FeSV‐transformed mink cells. Four positive hybrids were cloned twice in soft agar, established as stable lines, and grown in defined serum‐free medium to facilitate purification of homogeneous antibodies. The monoclonal antibodies were used to assay SM‐FeSV‐specific products by “immunoblotting” of elcctrophoretically separated proteins, and by fixed‐cell immunofluorescence.