title: Homogeneous assay for detection of active Epstein-Barr nuclear antigen 1 by thrombin activity modulation.
abstract: Epstein-Barr virus (EBV) has been associated with several malignancies as Burkitt's lymphoma, nasopharyngeal carcinoma, and Hodgkin's disease. In those diseases, Epstein-Barr nuclear antigen 1 (EBNA-1) is constitutively expressed. Here, we reported an innovative system to detect active EBNA-1 protein in a homogeneous assay. The system is based on the modulation of thrombin activity by a self-complementary single stranded DNA (scssDNA), which was designed and synthesized to mimic the palindromic target sites of EBNA-1 in the EBV genome. This model system showed a limit of detection of 3.75 ng mL(-1) of active EBNA-1 protein with a dynamic detection range from 3.75 to 250 ng mL(-1) with a correlation coefficient of 0.997. This new homogeneous assay for active EBNA-1 protein detection and quantification provides a very useful tool for rapid screening of EBNA-1 blockers in biomedical research.
authors: Gaizka
authors: G
authors: Garai-Ibabe
authors: Ruta
authors: R
authors: Grinyte
authors: Allon
authors: A
authors: Canaan
authors: Valeri
authors: V
authors: Pavlov
mesh_terms: D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019309:Epstein-Barr Virus Nuclear Antigens / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D013917:Thrombin / Q000737:chemistry / Q000378:metabolism*; D013997:Time Factors
chemical_list: D004277:DNA, Single-Stranded; D019309:Epstein-Barr Virus Nuclear Antigens; D013917:Thrombin; C417029:EBV-encoded nuclear antigen 1