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US 2008/0166762 A1Published application
HETEROLOGOUS PRODUCTION OF INTERLEUKIN 1 RECEPTOR ANTAGONIST (IL-1Ra) IN PICHIA PASTORIS
AssigneeReliance Life Science PVT.LTD.
InventorsLivy Shivraj, Pradnya Palande, Venkata Ramana Kondiboyina
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Abstract
The invention relates to the production and expression of Interleukin 1 receptor antagonist (IL-1Ra) in expression systems, such as Pichia Pastoris (P. pastoris). For example, the present invention encompasses an engineered Pichia strain x-33/pGAPZαA/IL-1Ra constructed by cloning the IL-1Ra gene in pGAPZαA and transforming the recombinant vector into Pichia Pastoris x-33.
Claims13
A method of producing a purified Interleukin-1 receptor antagonist (IL-1Ra) using a Pichia pastoris (P. pastoris) expression system, comprising the steps of:
- a) obtaining cDNA comprising a nucleic acid encoding an IL-1Ra protein, wherein the cDNA is prepared from RNA obtained from HS-5 cells;
- b) inserting the cDNA into a plasmid comprising a GAP promoter;
- c) transforming a P. pastoris host cell with the plasmid;
- d) expressing IL-1Ra protein in the host cell;
- and
- e) purifying the IL-1Ra protein;
- wherein the method involves continuous fermentation, and at least 10% of total protein secreted by the host cell into the fermentation medium is IL-1Ra protein.
4 more elements
Dependencies are inferred from the claim text.