Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Reexamination Certificate
2007-12-04
2007-12-04
Benzion, Gary (Department: 1637)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C435S091100, C435S091200
Reexamination Certificate
active
11234717
ABSTRACT:
To maximize the yield of protein from a baculovirus system, optimal infection of the host cell culture must be achieved; in order to obtain such optimal infection, the titer of the virus inoculation must be known. The present invention is the development of a simple, rapid, and universally applicable titration method that involves the direct detection of the viral DBP gene derived from AcNPV (AcNPV DBP) as a target for quantitative titer determination of baculovirus and the use of biotin specific probes directed to viral DBP gene. The procedure entails the amplification of the AcNPV DBP gene by using the PCR technique in the presence of digoxigenin-11-dUTP from the negative control (non-infected) and infected samples, and the synthesis of the specific biotin label nucleotide probes directed to the AcNPV DBP gene. These specific probes are then used in the Enzyme Linked Immunosorbent Assay (ELISA) using the immobilized streptavidin on polystyrene microtitration plates for the quantitative determination of baculovirus titer. The plot of O.D.λ=405 nmagainst the log of the titer (pfu/mL) generated a straight line. The linear range for titer determination of baculovirus was between 102and 105pfu/mL for 50 μL of supernatant.
Benzion Gary
Calamita Heather G.
Vista Biologicals Corporation
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