Two monoclonal antibodies (mAbs), 10-2 and 10-5, both directed against recombinant hirudin variant 2-Lys47 (rHV2), were selected for their high affinity and epitopic specificities to develop a two-site immunoassay of rHV2. The mAb concentrations, incubation time, and temperature were optimized. The immunoassay has a detection limit for rHV2 of 45 ng/L in plasma and 30 ng/L in urine. The reactiv...

BACKGROUND: For whole blood aggregation on the Multiplate® system, the use of thrombin inhibitor as anticoagulant is recommended. So far sample tubes containing liquid lepirudin were provided (Dynabyte, Munich, Germany). They are not sterile and have to be stored refrigerated. For better handling now also sterile tubes with dried hirudin, storable at room temperature, are also provided (Dynabyt...

Site-directed mutagenesis has been used to examine the importance of each of the acidic C-terminal residues of hirudin in the formation of its complex with alpha-thrombin. The contribution to binding energy of acidic residues in the 11 C-terminal amino acids varied from 2.3 to 5.9 kJ.mol-1. The differences between the contributions of individual residues were smaller than would be expected from...

OBJECTIVE To develop small-diameter vascular grafts with a microstructure similar to native matrix fibers and with chemically modified microfibers to prevent thrombosis. METHODS AND RESULTS Microfibrous vascular grafts (1-mm internal diameter) were fabricated by electrospinning, and hirudin was conjugated to the poly (L-lactic acid) microfibers through an intermediate linker of poly(ethylene ...

On former occasions we have discussed repeatedly the action of tissue extract and serum separately, as well as their combined action on fluoride, hirudin, and peptone plasma of vertebrate blood,l We pointed out the loss in power of a mixture of serum and tissue extract on standing and the difference in the behavior of fluoride plasma (and similar plasmas) in which coagulation had been prevented...

T he TIMI-9A1 and GUSTO-11A2 trial preliminary reports, in this issue of Circulation by highly experienced and respected investigators, show unexpectedly high rates of intracranial bleeds (ICBs) associated with administration of two plasminogen activators (streptokinase [SK] and tissue-type plasminogen activator [TPAI) and two intravenously administered anticoagulants (heparin and hirudin). Ano...

Neomycin was demonstrated to inhibit the binding of thrombin to intact human platelets. The effects of neomycin on both thrombin binding and thrombin-induced changes in inositol phospholipid metabolism could be reproduced by the thrombin antagonist hirudin. We propose that neomycin inhibits thrombin-induced platelet activation by interference with the cellular receptor.