Identification and molecular characterisation of proteins, expressed in the Ixodes ricinus salivary glands

A method of inhibiting thrombus formation in the blood of a human patient, the method comprising:

• (A) a method of inhibiting thrombus formation in the blood of a human patient wherein the blood contacts an exposed structure other than the patient's endothelium, the method comprising introducing to the patient's blood a sufficient amount to inhibit thrombus formation of an isolated fusion protein which comprises a polypeptide having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:36 fused to a heterologous polypeptide, wherein said polypeptide has a kunitz-type-protease-inhibitor (KPI) domain, wherein the KPI domain of the polypeptide comprises Phe at position corresponding to position 40 of SEQ ID NO:36, Gly at position corresponding to position 44 of SEQ ID NO:36;
• Cys at position corresponding to position 45 of SEQ ID NO:36, Phe at position corresponding to position 52 of SEQ ID NO:36, and Cys at position corresponding to position 58 of SEQ ID NO:36;
• or
• (B) a method of inhibiting thrombus formation, wherein the formation is inhibited after cutting the patient's vasculature, comprising cutting the patient's vasculature and introducing to the patient's blood flow a sufficient amount to inhibit thrombus formation of an isolated fusion protein which comprises a polypeptide having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:36 fused to a heterologous polypeptide, wherein said polypeptide has a kunitz-type-protease-inhibitor (KPI) domain, wherein the KPI domain of the polypeptide comprises Phe at position corresponding to position 40 of SEQ ID NO:36, Gly at position corresponding to position 44 of SEQ ID NO:36;
• Cys at position corresponding to position 45 of SEQ ID NO:36, Phe at position corresponding to position 52 of SEQ ID NO:36, and Cys at position corresponding to position 58 of SEQ ID NO:36.

A method of inhibiting the contact phase pathway, comprising:

• causing a human's blood to contact a sufficient amount to inhibit the contact phase pathway of an isolated fusion protein which comprises a polypeptide having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:36 fused to a heterologous polypeptide, wherein said polypeptide has a kunitz-type-protease-inhibitor (KPI) domain, wherein the KPI domain of the polypeptide comprises Phe at position corresponding to position 40 of SEQ ID NO:36, Gly at position corresponding to position 44 of SEQ ID NO:36;
• Cys at position corresponding to position 45 of SEQ ID NO:36, Phe at position corresponding to position 52 of SEQ ID NO:36, and Cys at position corresponding to position 58 of SEQ ID NO:36;
• and
• exposing the human's blood to a trigger selective for the contact phase pathway.

A method of preventing clot formation in the blood of a human patient without interfering with the bleeding time, the method comprising:

• (A) a method of preventing clot formation in the blood of a human patient without interfering with the bleeding time, wherein the blood contacts an exposed structure other than the patient's endothelium, the method comprising introducing to the patient's blood a sufficient amount to inhibit thrombus formation of an isolated fusion protein which comprises a polypeptide having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:36 fused to a heterologous polypeptide, wherein said polypeptide has a kunitz-type-protease-inhibitor (KPI) domain, wherein the KPI domain of the polypeptide comprises Phe at position corresponding to position 40 of SEQ ID NO:36, Gly at position corresponding to position 44 of SEQ ID NO:36;
• Cys at position corresponding to position 45 of SEQ ID NO:36, Phe at position corresponding to position 52 of SEQ ID NO:36, and Cys at position corresponding to position 58 of SEQ ID NO:36;
• or
• (B) a method of preventing clot formation in the blood of a human patient without interfering with the bleeding time, wherein the formation is inhibited after cutting the patient's vasculature, comprising cutting the patient's vasculature and introducing to the patient's blood flow a sufficient amount to inhibit thrombus formation of an isolated fusion protein which comprises a polypeptide having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:36 fused to a heterologous polypeptide, wherein said polypeptide has a kunitz-type-protease-inhibitor (KPI) domain, wherein the KPI domain of the polypeptide comprises Phe at position corresponding to position 40 of SEQ ID NO:36, Gly at position corresponding to position 44 of SEQ ID NO:36;
• Cys at position corresponding to position 45 of SEQ ID NO:36, Phe at position corresponding to position 52 of SEQ ID NO:36, and Cys at position corresponding to position 58 of SEQ ID NO:36.