title: A new fluorescent based screening system for high throughput screening of drugs targeting HBV-core and HBsAg interaction.
abstract: The existing screening systems for anti-hepatitis B virus (anti-HBV) drug discovery is time-consuming mainly due to the laborious detection system it is using. A new fluorescence based screening system for high throughput anti-HBV drug discovery was created by tagging hepatitis B surface antigen (HBsAg) with monomeric red fluorescent protein and hepatitis B virus (HBV) core protein with enhanced green fluorescent protein. The two constructs were co-transfected on to Hep3B cells and the transfection was stabilized by fluorescent activated cell sorter (FACS). The fusion proteins expressed through the secretory protein pathway as evidenced by localization with ER-Tracker and tubulin tracker. The new system has given analogues results like that of conventional enzyme-linked immunosorbent assay (ELISA). Hence it can be of very high potential for large scale drug screening systems.
authors.affiliation: Department of Botany, Govt. Victoria College, Palakkad, Kerala, India.
authors.forename: V
authors.initials: V
authors.lastname: Suresh
authors.affiliation: Department of Botany, Maharajas College, Ernakulam, Kerala, India.
authors.forename: K A
authors.initials: KA
authors.lastname: Krishnakumar
authors.affiliation: Plant Based Bioactives and Disease Biology Lab, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, Kerala, India. Electronic address: ashavv@rediffmail.com.
authors.forename: V V
authors.initials: VV
authors.lastname: Asha
mesh_terms: D000998:Antiviral Agents / Q000008:administration & dosage / Q000032:analysis* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D016503:Drug Delivery Systems / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005456:Fluorescent Dyes / Q000032:analysis*; D056945:Hep G2 Cells; D006514:Hepatitis B Surface Antigens / Q000032:analysis* / Q000378:metabolism; D006515:Hepatitis B virus / Q000737:chemistry* / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D014779:Virus Replication / Q000187:drug effects / Q000502:physiology
chemical_list: D000998:Antiviral Agents; D005456:Fluorescent Dyes; D006514:Hepatitis B Surface Antigens
keywords: Cell based assay system; Fluorescence; HBV-core; HBsAg; Lamivudine; eGFP; mRFP