Monoclonal antibodies to the light chain region of human factor XII and methods of preparing and using the same
AssigneeTemple University of the Commonwealth System of Higher Education
InventorRobin A. Pixley
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Abstract
Novel cell lines produce monoclonal antibody to the light chain region of human factor XII. Three such cell lines, ATCC #HB-9703 through ATCC #HB-9704, are formed by fusing SP2/0-Ag14 myeloma cells with spleen cells from BALB/c AnSkh mice immunized with human factor XIIf. Biochemical and therapeutic uses of the monoclonal antibodies are provided.
Claims27
A monoclonal antibody which specifically binds to an antigenic determinant in the light chain region of human factor XII, which determinant is preserved and present in factor XIIa and factor XIIf, said monoclonal antibody having the ability to inhibit the coagulant activity of factor XIIa and the enzyme activity of factor XIIa and factor XIIf.
A method for producing a monoclonal antibody which binds to an antigenic determinant of the light chain region of human factor XII, which determinant is preserved and present in factor XIIa and factor XIIf, said monoclonal antibody having the ability to inhibit the coagulant activity of factor XIIa and the enzyme activity of factor XIIa and factor XIIf, which method comprises the steps of:
- (a) immunizing a splenocyte donor with an immunogen containing the light chain region of human factor XII;
- (b) removing the spleen from said donor and making a suspension of the spleen cells;
- (c) fusing said spleen cells with myeloma cells;
- (d) diluting and culturing the fused cells in separate wells in a medium which will not support the unfused myeloma cells;
- (e) assaying the supernatant in each well containing a hybridoma for the presence of antibody to the light chain region of human factor XII;
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- (f) selecting and cloning a hybridoma secreting antibody which binds to an antigenic determinant in the light chain region of human factor XII, which determinant is preserved and present in factor XIIa and factor XIIf, said monoclonal antibody having the ability to inhibit the coagulant activity of factor XII and the enzyme activity of factor XIIa and factor XIIf.
A method for producing a monoclonal antibody against the light chain region of human factor XII which comprises culturing a hybridoma selected from the group consisting of hybridomas ATCC #HB-9703, ATCC #HB-9704 and ATCC #HB-9705, and recovering the secreted monoclonal antibodies from the culture medium.
A method for producing a monoclonal antibody against the light chain region of human factor XII which comprises the steps of injecting into a mouse a hybridoma selected from the group consisting of hybridomas ATCC #HB-9703, ATCC #HB-9704, and ATCC #HB-9705, and recovering the secreted monoclonal antibodies form the mouse ascitic fluid or serum.
A composition comprising a continuous cell line producing monoclonal antibody which specifically binds to an antigenic determinant in the light chain region of human factor XII, which determinant is preserved and present in factor XIIa and factor XIIf, comprising a cell hybrid of a myeloma fused to a spleen cell from a donor previously immunized with an immunogen containing human factor XII light chain, and a culture medium for said hybrid, said monoclonal antibody produced by said hybrid having the ability to inhibit the coagulant activity of factor XIIa and the enzyme activity of factor XIIa and factor XIIf.
The cell line ATCC #HB-9703.
The cell line ATCC #HB-9704.
The cell line ATCC #HB-9705.
A method for removing human factor XII and enzymatically active factor XII fragments from a liquid with an immobilized monoclonal antibody which binds to an antigenic determinant in the light chain region of human factor XII, which determinant is preserved and present in such enzymatically active factor XII fragments, and absorbing said factor XII and factor XII fragments from the liquid.
Dependencies are inferred from the claim text.