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Method of increasing the hydrodynamic volume of polypeptides by attaching to gonadotrophin carboxy terminal peptides

  • US 9,808,534 B2
  • Filed: 11/20/2013
  • Issued: 11/07/2017
  • Est. Priority Date: 11/20/2012
  • Status: Active Grant
First Claim
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1. 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.

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