A competition immunoassay for characterizing the reverse transcriptase of mammalian RNA tumor viruses

Amos Panet*, Zipora Kra-oz

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

A competition enzyme immunoassay for reverse transcriptase has been developed. This assay is based on competition between the enzymatically active reverse transcriptase probe and a sample tested for the presence of reverse transcriptase antigen on a limited amount of monospecific antibody. The extent of competition is determined from the residual DNA polymerase activity left in the probe. Any reverse transcriptase activity in the test sample that may interfere with the enzymatic assay is irreversibly inhibited by pretreatment with N-ethylmaleimide. Thus, increasing amounts of test antigen neutralize more antibodies and as a result restore more of the reverse transcriptase probe activity. By means of the competition immunoassay we were able to show type, group, and interspecies antigenic determinants in the reverse transcriptase of mammalian RNA tumor viruses. The competition immunoassay has also proved useful for quantitating the amount of reverse transcriptase present in virions. The enzyme was found to comprise 2.5% of the total proteins in Moloney murine leukemia virus.

Original languageEnglish
Pages (from-to)95-101
Number of pages7
JournalVirology
Volume89
Issue number1
DOIs
StatePublished - Aug 1978

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