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PMID: 6312092 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Isolation of monoclonal antibodies that recognize the transforming proteins of avian sarcoma viruses.

Journal of virology ·Vol. 48 ·No. 2 ·1983-11-00 ·Pages 352-60

Lipsich LA, Lewis AJ, Brugge JS

Abstract

Thirteen clones of hybrid cells which synthesize antibodies directed against the Rous sarcoma virus (RSV) transforming protein, pp60src, were isolated. Mouse myeloma cells were fused with spleen cells from mice that had been immunized with purified pp60src from bacterial recombinants which direct the synthesis of the RSV src gene. The hybridomas which survived the selection medium were screened by immunoprecipitation of pp60src from 32P-labeled lysates of RSV-transformed cells. Monoclonal antibodies produced by subclones derived from 13 hybridomas recognized pp60src encoded by the Schmidt-Ruppin and Prague strains of RSV and the cellular homolog of pp60src. Antibody from clone 261 had a high affinity for the viral yes gene product, and antibodies from clones 443 and 463 recognized the transforming proteins encoded by viruses containing the related transforming genes fps and ros. Several other clones had a low affinity for the viral yes, fps, and ros gene products which could be detected by in vitro phosphorylation of the transforming proteins after immunoprecipitation with the monoclonal antibody. All of the monoclonal antibodies allowed phosphorylation of pp60src and casein in an immune complex-bound reaction.

MeSH Terms
Animals Antibodies, Monoclonal/isolation & purification Antibody Affinity Antigens, Polyomavirus Transforming Avian Sarcoma Viruses/analysis Caseins/metabolism Cross Reactions Hybridomas Mice Oncogene Protein pp60(v-src) Phosphorylation Protein Kinases/metabolism Viral Proteins/immunology,metabolism
Chemicals
Antibodies, Monoclonal Antigens, Polyomavirus Transforming Caseins Viral Proteins Protein Kinases Oncogene Protein pp60(v-src)
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Lipsich L A
Lewis A J
Brugge J S
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37 references, click to expand
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.48.2.352-360.1983
Published
1983-11-00
Pages
352-60
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC255359
Subset
IM
Grants
NCI NIH HHS · CA 27951 · United States
External Links
PubMed source DOI: 10.1128/JVI.48.2.352-360.1983 PMC Full Text View journal details
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