The Use of Antibodies against Synthetic Peptides for Studying the EBV Nuclear Antigen

Abstract

Five peptides corresponding to amino acid sequences predicted from the BamHI K fragment of the EBV genome have been synthesized (Table 1). The antisera raised against peptide no. 107, a copolymer of alanine and glycine deduced from the third internal repeat (IR3) sequence, gave brilliant nuclear staining in the anticomplement immunof1ourescence assay (AC IF) on eight EBV-carrying lines (Figure 1a),whereas five EBV-negative lines were not stained (Table 2). The nuclear staining was competed out by addition of the synthetic peptide (Figure 1b). 19 out of 21 EBNA-positive sera reacted with the synthetic peptide in an ELISA test, whereas EBV- negative sera did not react or gave very weak reactions (Figure 2). The 19 EBNA-positive sera that reacted with the peptide were all healthy donor sera whereas the 3 EBNA-positive sera that failed to react were Burkitt lymphoma (BL) patient sera.In a later series of experiments 34 EBNA-positive healthy donor sera were all found to react with the synthetic peptide, whereas out of 48 BL sera 19 did not have antibodies to this peptide (not shown).