Preparation of human papillomavirus E1 having helicase activity and method therefor
AssigneeBoehringer Ingelheim (Canada) Ltd.
InventorsAlex Pelletier, Chris M. Farnet
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Abstract
The present invention relates to a method for isolating cloned papillomavirus E1 protein from a eukaryotic expression system having demonstrable and reproducible viral helicase activity and preparation containing essentially pure E1 protein. The invention further relates to the use of this novel E1 protein preparation in a screening assay for identifying antiviral agents. More particularly a high throughput assay to screen for agents capable of inhibiting HPV DNA replication. The assay is based on measuring the effect of antiviral agents on the activity of the E1 protein and more specifically on its helicase activity.
Claims23
A method for isolating recombinant papillomavirus E1 protein having quantifiable unwinding activity comprising the steps of:
- producing an E1 recombinant protein in a eukaryotic expression system and isolating a nuclei preparation thereof;
- and
- extracting E1 recombinant protein from said nuclei preparation in a buffer comprising salt at a concentration equal to or below isotonic concentration.
Dependencies are inferred from the claim text.