Preparation of monoclonal antibodies to the thrombin/hirudin complex.
Monoclonal antibodies (mAbs) binding to the thrombin/hirudin (T.H) complex were prepared by either immunizing mice with hirudin and by screening for the mAbs cross-reacting with the T.H complex (group I), or by immunizing the animals directly with the T.H complex (group II). Epitope mapping of the mAbs of group I indicated that all the mAbs were binding to the hirudin N-terminal core domain only (residues 1-43). Among the mAbs raised against the T.H complex (group II), one mAb recognized an antigenic determinant expressed selectively upon binding of hirudin to thrombin. A double antibody sandwich type ELISA combining the mAb of group II with a mAb of group I was developed, allowing the determination of the T.H complex in human citrated plasma down to nanogram concentration levels.
Schlaeppi
J M
JM
Pharmaceuticals Research Laboratories, Ciba-Geigy Limited, Basle, Switzerland.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000918:Antibody Specificity; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D006629:Hirudins / Q000097:blood / Q000276:immunology*; D051379:Mice; D010446:Peptide Fragments / Q000276:immunology; D013917:Thrombin / Q000032:analysis / Q000276:immunology*
D000911:Antibodies, Monoclonal; D006629:Hirudins; D010446:Peptide Fragments; D013917:Thrombin