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US 9,808,534 B2Granted
Method of increasing the hydrodynamic volume of polypeptides by attaching to gonadotrophin carboxy terminal peptides
AssigneeOPKO Biologics Ltd.
InventorsOren Hershkovitz, Ahuva Bar-Ilan
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Abstract
This invention is directed to the use of a chorionic gonadotrophin carboxy terminal peptide (CTP) or fragments thereof for modifying a polypeptide or a fragment thereof in order to increase the hydrodynamic volume of the polypeptide or fragment thereof.
Claims8
A method of incrementally increasing the hydrodynamic size of a recombinant polypeptide of interest wherein said recombinant polypeptide of interest is selected from the group consisting of a coagulation factor IX (FIX) and a coagulation factor VIIa (FVIIa),
- said method comprising a step of recombinantly fusing
- (a) three chorionic gonadotrophin carboxy terminal peptide CTP units to the carboxy terminus of said coagulation factor IX (FIX) and no CTPs attached to the N-terminus;
- or
- (b) three CTP units to the carboxy terminus of said coagulation factor VIIa (FVIIa) and no CTPs attached to the N-terminus;
- followed by a step of expressing the recombinant CTP-modified polypeptides in a Chinese hamster ovary (CHO) host cell, wherein expression comprises glycosylating said CTP-units and the glycosylation comprises O-glycosylation;
- wherein said glycosylated CTP units incrementally increase the hydrodynamic size of the FIX by an increment of about 48-53 kDA per each of said glycosylated CTP units, and,
- wherein said glycoslyated CTP units incrementally increase the hydrodynamic size of the FVIIa by an increment of about 43-50 kDA per each of said glycosylated CTP units,
- thereby incrementally increasing the hydrodynamic size of said recombinant polypeptide of interest.
5 more elements
Dependencies are inferred from the claim text.