Granted Patent B2

US 8,101,394 · App. 12/744,376 · Granted Jan 24, 2012

Abstract

Claims (15)

1. A polynucleotide comprising a nucleotide sequence encoding a polypeptide having:

(a) a single N-terminal kringle domain, wherein the last four amino acid residues within the single N-terminal kringle domain are identical to residues at positions 91-94 of SEQ ID NO: 2; and

(b) a C-terminal domain activation site and serine protease domain;

wherein the polypeptide binds to immobilized lysine; and

wherein the encoded polypeptide is at least 95% identical to the sequence shown in SEQ ID NO:2.

2. The polynucleotide of claim 1 , wherein the encoded polypeptide comprises the sequence shown in SEQ ID NO: 2.

3. The polynucleotide of claim 1 , wherein the nucleotide sequence of the polynucleotide is the sequence shown in SEQ ID NO: 1, or a degenerate variant thereof.

4. The polynucleotide of claim 1 , wherein the encoded polypeptide exhibits a lower binding affinity for fibrinogen than the binding affinity for fibrinogen of mini-plasmin.

5. The polynucleotide of claim 1 , wherein the encoded polypeptide exhibits higher binding affinity for partially cleaved fibrin than the binding affinity for partially cleaved fibrin of mini-plasmin.

6. The polynucleotide of claim 1 , wherein the encoded polypeptide has reduced immunogenicity as compared to a reference polypeptide, wherein the reference polypeptide has a primary amino acid sequence identical to the primary amino acid sequence of the polypeptide with the proviso that the last four amino acid residues of the single N-terminal kringle domain of the reference polypeptide are not identical to residues at positions 91-94 of SEQ ID NO:2.

7. The polynucleotide of claim 1 , wherein amino acid residues 2-9 of the encoded polypeptide are identical to residues at positions 2-9 of SEQ ID: NO: 2.

8. The polynucleotide of claim 1 , wherein the encoded polypeptide is at least 98% identical to the sequence shown in SEQ ID NO: 2.

9. A vector comprising the polynucleotide of claim 1 .

10. A cell comprising the vector of claim 9 .

11. A method of producing a polypeptide comprising expressing the polynucleotide in the cell of claim 10 .

Assignments (27)