title: Label free inhibitor screening of hepatitis C virus (HCV) NS5B viral protein using RNA oligonucleotide.
abstract: Globally, over 170 million people (ca. 3% of the World's population) are infected with the hepatitis C virus (HCV), which can cause serious liver diseases such as chronic hepatitis, evolving into subsequent health problems. Driven by the need to detect the presence of HCV, as an essential factor in diagnostic medicine, the monitoring of viral protein has been of great interest in developing simple and reliable HCV detection methods. Despite considerable advances in viral protein detection as an HCV disease marker, the current enzyme linked immunosorbent assay (ELISA) based detection methods using antibody treatment have several drawbacks. To overcome this bottleneck, an RNA aptamer become to be emerged as an antibody substitute in the application of biosensor for detection of viral protein. In this study, we demonstrated a streptavidin-biotin conjugation method, namely, the RNA aptamer sensor system that can quantify viral protein with detection level of 700 pg mL(-1) using a biotinylated RNA oligonucleotide on an Octet optical biosensor. Also, we showed this method can be used to screen inhibitors of viral protein rapidly and simply on a biotinylated RNA oligonucleotide biosensor. Among the inhibitors screened, (-)-Epigallocatechin gallate showed high binding inhibition effect on HCV NS5B viral protein. The proposed method can be considered a real-time monitoring method for inhibitor screening of HCV viral protein and is expected to be applicable to other types of diseases.
authors.affiliation: Radiation Research Division for Biotechnology, Korea Atomic Energy Research Institute, 1266 Shinjeong-dong, Jeongeup-si, Jeollabuk-do 580-185, Korea. chroh@kaeri.re.kr
authors.forename: Changhyun
authors.initials: C
authors.lastname: Roh
authors.forename: Sang Eun
authors.initials: SE
authors.lastname: Kim
authors.forename: Sung-Kee
authors.initials: SK
authors.lastname: Jo
mesh_terms: D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology; D052157:Aptamers, Nucleotide / Q000737:chemistry*; D001710:Biotin / Q000737:chemistry; D002392:Catechin / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology; D003199:Computer Systems; D016174:Hepacivirus / Q000302:isolation & purification*; D006526:Hepatitis C / Q000175:diagnosis*; D006801:Humans; D019809:Streptavidin / Q000737:chemistry; D017361:Viral Nonstructural Proteins / Q000032:analysis* / Q000037:antagonists & inhibitors; D012324:RNA-Dependent RNA Polymerase
chemical_list: D000998:Antiviral Agents; D052157:Aptamers, Nucleotide; D001710:Biotin; D002392:Catechin; D019809:Streptavidin; D017361:Viral Nonstructural Proteins; C045651:epigallocatechin gallate; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase