Conformation-specific anti-von Willebrand factor antibodies
AssigneeThe Brigham and Women's Hospital, Inc.
InventorsRobert I. Handin, Huabing Yuan, Anne McLeod
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Abstract
The present invention is directed to antibodies and antibody fragments that bind specifically to the active conformation of human von Willebrand factor. Most preferred are recombinantly produced single chain variable immunoglobulin fragments. Because the antibodies or antibody fragments act only at the sites of thrombus formation and do not interfere with the normal activity of circulating platelets, they are particularly well suited for use as antithrombotic agents in a wide variety of applications.
Claims17
A composition comprising an antibody that binds selectively to the active conformation of human von Willebrand Factor (vWF) and wherein said antibody inhibits the binding of vWF to platelets.
A method of identifying a single chain variable region immunoglobulin (ScFv) fragment that binds selectively to the active conformation of vWF, comprising:
- a) immunizing an animal with an immunogen derived from the A1 region of human vWF;
- b) isolating mRNA from the immunized animal of step a);
- c) producing a cDNA library from the isolated mRNA of step b) in a bacteriophage capable of displaying ScFv fragments encoded by the cloned cDNA;
- and
- d) screening the library of step c) to identify a bacteriophage displaying a ScFv fragment that binds selectively to the active conformation of vWF.
The method of making and ScFv fragment that binds selectively to the active conformation of vWF, comprising:
- a) immunizing an animal with an immunogen derived from the A1 region of human vWF;
- b) isolating mRNA from the immunized animal of step a);
- c) producing a cDNA library from the isolated mRNA of step b) in a bacteriophage capable of displaying ScFv fragments encoded by the cloned cDNA;
- and
- d) screening the library of step c) to identify a bacteriophage displaying a ScFv fragment that binds selectively to the active conformation of vWF.
- e) subcloning the recombinant DNA encoding the ScFv fragment identified in step d) in an expression vector;
- f) transforming a host cell capable of expressing recombinant ScFv with the expression vector of step e);
- and
- g) purifying the recombinant ScFv produced by the host cell of step f).
A method of inhibiting the binding of human vWF to platelets, comprising contacting said human vWF with an antibody that binds selectively to the active conformation of vWF.
A method of preventing thrombus formation in a patient, comprising administering a pharmaceutical composition comprising an antibody that binds selectively to the active conformation of human vWF and that thereby prevents the binding of vWF to platelets, wherein said pharmaceutical composition is administered at a dosage sufficient to inhibit said thrombus formation.
Dependencies are inferred from the claim text.