Identification of a purified complement fixing antigen as the Epstein Barr virus determined nuclear antigen (EBNA) by its binding to metaphase chromosomes

S. Ohno, J. Luka, T. Lindahl, G. Klein

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Abstract

A soluble complement-fixing antigen carried by Epstein-Barr virus (EBV)-transformed human cells has been previously extracted from cell nuclei and purified by DNA-cellulose chromatography. On addition of this antigen the methanol/acetic acid-fixed metaphase chromosomes, followed by exposure to human sera containing antibodies against the EBV-determined nuclear antigen (EBNA), brilliant positive staining was obtained by anti-complement immunofluorescence. There was no staining after exposure to EBV-negative sera. Moreover, a nuclear protein fraction, prepared from an EBV-negative cell line in an analogous fashion, failed to induce the staining reaction. These data identify the soluble purified antigen as the EBV-determined nuclear antigen. The purified antigen has a molecular weight of 174,000 ± 15,000, as determined by sucrose gradient centrifugation and gel filtration experiments. In neutral buffers containing 0.5-1.0 M NaCl, the antigen dissociates into a form of approximately one-half the original molecular weight with retained complement-fixing activity. This 'monomer' has a molecular weight of 98,000 ± 8,000.

Original languageEnglish
Pages (from-to)1605-1609
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume74
Issue number4
DOIs
StatePublished - 1977
Externally publishedYes

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