American Society of Hematology

Figure 5

$Figure 5. Enhanced coagulation activity of rFVIII-PEG-Lip is removed from PPP by ultracentrifugation. HemA mice were dosed with 8 IU/kg rFVIII (n = 20) or rFVIII-PEG-Lip (n = 20). At 5 minutes after dosing, blood was collected from the vena cava and tested individually by ROTEM. (A) Whole blood (3 μL) was added to 300 μL blood collected from naive HemA mouse. (B) Blood from each mouse was centrifuged at 300g for 5 minutes and 3 μL of the PRP fraction (supernatant) was added to 300 μL freshly collected naive HemA mouse blood. (C) Immediately after the collection of the 3 μL PRP from the low-speed spin, the blood from each mouse was further centrifuged at 1500g for 5 minutes. Three microliters from the PPP fraction (supernatant) was added to 300 μL freshly collected naive HemA mouse blood. (D) Thawed plasma was subjected to ultracentrifugation at 100000g for 20 minutes and 3 μL of the supernatant was added to 300 μL freshly collected naive HemA mouse blood. Two-tailed P values were determined by Mann-Whitney test. P > .05 for PPP versus UFP from rFVIII-treated animals; P = .016 for PPP versus UFP from rFVIII-PEG-Lip–treated animals; P = .008 for PPP from rFVIII- versus rFVIII-PEG-Lip–treated animals; and P > .05 for UFP from rFVIII- versus rFVIII-PEG-Lip–treated animals.$

Enhanced coagulation activity of rFVIII-PEG-Lip is removed from PPP by ultracentrifugation. HemA mice were dosed with 8 IU/kg rFVIII (n = 20) or rFVIII-PEG-Lip (n = 20). At 5 minutes after dosing, blood was collected from the vena cava and tested individually by ROTEM. (A) Whole blood (3 μL) was added to 300 μL blood collected from naive HemA mouse. (B) Blood from each mouse was centrifuged at 300g for 5 minutes and 3 μL of the PRP fraction (supernatant) was added to 300 μL freshly collected naive HemA mouse blood. (C) Immediately after the collection of the 3 μL PRP from the low-speed spin, the blood from each mouse was further centrifuged at 1500g for 5 minutes. Three microliters from the PPP fraction (supernatant) was added to 300 μL freshly collected naive HemA mouse blood. (D) Thawed plasma was subjected to ultracentrifugation at 100000g for 20 minutes and 3 μL of the supernatant was added to 300 μL freshly collected naive HemA mouse blood. Two-tailed P values were determined by Mann-Whitney test. P > .05 for PPP versus UFP from rFVIII-treated animals; P = .016 for PPP versus UFP from rFVIII-PEG-Lip–treated animals; P = .008 for PPP from rFVIII- versus rFVIII-PEG-Lip–treated animals; and P > .05 for UFP from rFVIII- versus rFVIII-PEG-Lip–treated animals.

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