Detection of human serum antibodies to the BFRF3 Epstein-Barr virus capsid component by means of a DNA-binding assay.
A novel assay for antibodies to an immunodominant component of the Epstein-Barr virus (EBV) capsid antigen (VCA) complex was developed by creation of a chimeric protein containing the DNA-binding domain of the yeast GAL4 protein fused to the capsid antigen encoded by the BFRF3 gene of EBV. GAL4-BFRF3 antigen fusion protein bound specifically to a duplex DNA oligonucleotide containing GAL4-binding sites. Antibodies to the antigen were revealed by retardation of the electrophoretic mobility of the DNA-protein complex. Antibodies to the BFRF3 component of VCA became detectable approximately 2 months after onset of infectious mononucleosis. Kinetics of the antibody response to BFRF3 were identical using supershift or immunoblotting assays. Concordance between the DNA-binding assay and the classical indirect immunofluorescence assay for antibody to VCA was 97%. The GAL4 epitope assay is applicable for detection of antibodies to many cloned gene products.
Shedd
D
D
Department of Pediatrics, Yale University School of Medicine, New Haven, Connecticut 06520-8064, USA.
Angeloni
A
A
Niederman
J
J
Miller
G
G
D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000276:immunology*; D001483:Base Sequence; D002213:Capsid / Q000276:immunology*; D036022:Capsid Proteins; D003289:Convalescence; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins; D019084:Fluorescent Antibody Technique, Indirect; D005656:Fungal Proteins / Q000276:immunology; D006566:Herpesviridae Infections / Q000097:blood / Q000276:immunology*; D004854:Herpesvirus 4, Human / Q000276:immunology*; D006801:Humans; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D011993:Recombinant Fusion Proteins / Q000276:immunology; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors
D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D004268:DNA-Binding Proteins; C041788:Epstein-Barr viral capsid antigen; D005656:Fungal Proteins; C052130:GAL4 protein, S cerevisiae; D015345:Oligonucleotide Probes; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D004247:DNA