Wnt compositions and methods of process from serum-free culturing conditions
AssigneeAnkasa Regenerative Therapeutics, Inc.
InventorsPing Yuan, JR., Jill Helms, Ying Zhu, Bo Liu, Stefanie Gaster
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Abstract
Disclosed herein are methods and compositions for producing a Wnt polypeptide under a serum-free condition. Also disclosed herein are methods of purifying the Wnt polypeptide from a serum-free condition.
Claims26
A method of preparing a functionally active Wnt polypeptide, comprising:
- (a) co-expressing a Wnt polypeptide and a chaperone in a cell in a conditioned media to generate a plurality of Wnt polypeptide-chaperone complexes;
- (b) harvesting the plurality of Wnt polypeptide-chaperone complexes from the conditioned media;
- (c) incubating the plurality of Wnt polypeptide-chaperone complexes with a buffer comprising a sugar detergent to generate a mixture comprising a first Wnt composition comprising a functionally inactive Wnt polypeptide and a chaperone composition;
- (d) separating the first Wnt composition from the mixture with a column immobilized with a sulfonated polyaromatic compound to generate a second Wnt composition comprising the functionally active Wnt polypeptide and the sugar detergent;
- and
- (e) contacting the second Wnt composition with an aqueous solution of liposomes to generate a final Wnt composition comprising a functionally active Wnt polypeptide.
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A mammalian cell comprising an exogenous Wnt polypeptide and an exogenous chaperone protein.
A pharmaceutical composition, wherein the pharmaceutical composition comprises a liposomal Wnt3A polypeptide, wherein the amount of active Wnt3A polypeptide present in the pharmaceutical composition provides for an EC50 of about equal to or less than 0.5 ng/uL when administered to a bone graft material.
Dependencies are inferred from the claim text.