Example 9
A serial dilution of anti-ATβH hmAbs was made in hemA plasma starting at 250 nM with 0.1 U/mL of heparin. The mixture of antibody, plasma and heparin was incubated at room temperature for 20-30 min. Then 50 uL of this mixture was added to a 50 uL of diluted Innovin (1/2000) (Dade Behring), incubated for 4 min at 37° C., followed by adding 50 uL of 25 mM CaCl2 (HemSil). dPT test program was set on ACL Top coagulometer with acquisition time of 360 seconds. For dTP in AT deficient plasma, AT-DP was spiked in with either ATα or ATβ at a final concentration of 0.2 uM with 0.1 U/ml of heparin. Anti-ATβH mAbs was added to AT-DP/heparin/ATα or AT-DP/heparin/ATβ mixtures at a final concentration of 0.25 uM and incubated at room temperature for 20-30 min. For each reaction, a 50 uL of plasma/antibody/heparin mixture was added to 50 uL of diluted Innovin (1/4000), incubate 4 min at 37° C., followed by adding 50 uL of 25 mM CaCl2 (HemSil) as above.