AN IN VITRO ACTIVITY ASSAY FOR HUMAN HEPATITIS B VIRUS (HBV) DNA POLYMERASE, AND ITS USE FOR SCREENING INHIBITORS OF HBV DNA POLYMERASE

• Main Authors: LEONG, AI LIN, CON, CHONG JIN, LIM, GEK KEOW, CHEN, WEI NING
• Format: Patent
• Language: English
• Published: 19.07.2005
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PHYSICS; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; TESTING; VINEGAR; WINE

The present invention provides an in vitro activity assay for human hepatitis B virus (HBV) DNA polymerase, which comprises using, as the 5' oligonucleotide in PCR amplification of HBV DNA polymerase from a sample, an oligonucleotide into which has been incorporated the SP6 viral polymerase promoter, directly transcribing and translating the PCR products in the presence of a radio- labelled agent and measuring the priming of the HBV DNA polymerase. The present invention also provides the use of such an assay to assay activity of various serum samples, to screen for inhibitors of the HBV DNA polymerase and to test and/or screen potential anti- HBV drugs for their ability to inhibit DNA priming activity of human HBV DNA polymerase. Dati pronalazak obezbedjuje ogled za odredjivanje in vitro aktivnosti DNA polimeraze humanog virusa hepatitisa B (HBV), koji obuhvata korišćenje 5'-oligonukleotida 5' oligonukleotida kao prajmera za umnožavanje HBV DNA polimeraze PCR reakcijom iz uzorka, oligonukleotida u koji je umetnut promoter SP6 virusne polimeraze, direktno transkribujući i prepisujući PCR proizvode u prisustvu radioaktivno obeleženog sredstva i merenje (polimerizujuće) aktivnosti HBV DNA polimeraze. Ovaj pronalazak takodje obezbedjuje upotrebu takvog ogleda da bi se ispitala aktivnost različitih uzoraka seruma, za odredjivanje inhibitora HBV DNA polimeraze i za testiranje /ili odredjivanje potencijalnih lekova protiv HBV za njiihovu sposobnost da inhibiraju (polimerizujuće) aktivnosti DNA za HBV DNA polimerazu.[The present invention provides an in vitro activity assay for human hepatitis B virus (HBV) DNA polymerase, which comprises using, as the 5' oligonucleotide in PCR amplification of HBV DNA polymerase from a sample, an oligonucleotide into which has been incorporated the SP6 viral polymerase promoter, directly transcribing and translating the PCR products in the presence of a radio- labelled agent and measuring the priming of the HBV DNA polymerase. The present invention also provides the use of such an assay to assay activity of various serum samples, to screen for inhibitors of the HBV DNA polymerase and to test and/or screen potential anti- HBV drugs for their ability to inhibit DNA priming activity of human HBV DNA polymerase.