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 similar to 2 months after onset of infectious mononucleosis. Kinetics of the antibody response to BFRFS 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.
Detection of human serum antibodies to the BFRF3 Epstein-Barr virus capsid component by means of a DNA-binding assay / Shedd, D; Angeloni, Antonio; Niederman, J; Miller, G.. - In: THE JOURNAL OF INFECTIOUS DISEASES. - ISSN 0022-1899. - STAMPA. - 172:(1995), pp. 1367-1370. [10.1093/infdis/172.5.1367]
Detection of human serum antibodies to the BFRF3 Epstein-Barr virus capsid component by means of a DNA-binding assay.
ANGELONI, Antonio;
1995
Abstract
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 similar to 2 months after onset of infectious mononucleosis. Kinetics of the antibody response to BFRFS 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.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
