Genetically modified cells and methods for expressing recombinant heparanase and methods of purifying same
First Claim
1. A method of activating a mammalian heparanase protein, the method comprising digesting the mammalian heparanase protein with a protease capable of cleaving said mammalian heparanase protein at a region containing its most hydrophilic sites present within the first 170 N terminal amino acids of said mammalian heparanase protein, as determined using the Kyte-Doolittle method for calculating hydrophilicity, using the Wisconsin University GCG DNA analysis software, so as to release a catalytically active portion of said mammalian heparanase.
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Abstract
Bacterial, yeast and animal cells and methods for overexpressing recombinant heparanase in cellular systems, methods of purifying recombinant heparanase therefrom and modified heparanase species which serve as precursors for generating highly active heparanase by proteolysis.
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Citations
8 Claims
- 1. A method of activating a mammalian heparanase protein, the method comprising digesting the mammalian heparanase protein with a protease capable of cleaving said mammalian heparanase protein at a region containing its most hydrophilic sites present within the first 170 N terminal amino acids of said mammalian heparanase protein, as determined using the Kyte-Doolittle method for calculating hydrophilicity, using the Wisconsin University GCG DNA analysis software, so as to release a catalytically active portion of said mammalian heparanase.
- 5. A method of activating a modified mammalian heparanase protein, wherein the modified mammalian heparanase protein comprises an introduced protease cleavage recognition sequence at a region containing the most hydrophilic sites present within the first 170 N terminal amino acids of a natural mammalian heparanase protein as determined using the Kyte-Doolittle method for calculating hydrophilicity, using the Wisconsin University GCG DNA analysis software, the method comprising digesting the modified mammalian heparanase protein with a matching protease being capable of cleaving said introduced protease cleavage recognition sequence.
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