Purification and isolation of recombinant oxalate degrading enzymes and spray-dried particles containing oxalate degrading enzymes
AssigneeOxthera Intellectual Property AB
InventorsHarmeet Sidhu, Aaron Blake Cowley, Carl-Gustaf Golander, Qingshan Li
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Abstract
The present invention comprises methods and compositions for the reduction of oxalate in humans, and methods for the purification and isolation of recombinant oxalate reducing enzyme proteins. The invention provides methods and compositions for the delivery of oxalate-reducing enzymes in particle compositions. The compositions of the present invention are suitable in methods of treatment or prevention of oxalate-related conditions.
Claims20
Spray-dried particles comprising one or more isolated recombinant oxalate decarboxylase proteins encoded by the nucleotide sequence of SEQ ID NO:18, which are insoluble in the cytoplasm of a host cell and are not found as an inclusion body, and a polymeric material, wherein the activity of the one or more recombinant proteins at the most decreases to about 30% when incubated in a 3.2 mg/ml pepsin solution having a pH of about 3.2 for 40 minutes with the initial activity being set to 100%.
A method for isolating one or more recombinant oxalate decarboxylase proteins encoded by the nucleotide sequence of SEQ ID NO:18, that are insoluble in the cytoplasm of a host cell and are not found as an inclusion body, comprising:
- (a) separating one or more insoluble recombinant proteins not found as an inclusion body from soluble host cell proteins;
- and
- (b) solubilising the separated one or more recombinant proteins,
- wherein the activity of the one or more recombinant proteins at the most decreases to about 30% when incubated in a 3.2 mg/ml pepsin solution having a pH of about 3.2 for 40 minutes with the initial activity being set to 100%.
Dependencies are inferred from the claim text.