Comparison of FIXa binding to FVIIIa within Xase complex and FX interaction with Xase complex assembled with FVIIIa on synthetic phospholipids (n = 6, Mean ± SD) [10].

<p>Comparison of FIXa binding to FVIIIa within Xase complex and FX interaction with Xase complex assembled with FVIIIa on synthetic phospholipids (n = 6, Mean ± SD) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0113600#pone.0113600-Peters1" target="_blank">[10]</a>.</p

Thrombin generation profile of FVIII variants.

<p>Activity determination was based on equal chromogenic activity. Representative curves with selected concentrations are shown in (A) 1 IU/mL (B) 0.25 IU/mL (C) 0.0625 IU/mL. Select parameters are shown in (D) as peak thrombin and (E) as ETP.</p

Affinities of SC rFVIIIFc and rFVIIIFc for VWF by SPR (n = 6, Mean ± SD) [10].

<p>Affinities of SC rFVIIIFc and rFVIIIFc for VWF by SPR (n = 6, Mean ± SD) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0113600#pone.0113600-Peters1" target="_blank">[10]</a>.</p

Summary of PK parameters for SC rFVIIIFc and rFVIIIFc.

<p>Summary of PK parameters for SC rFVIIIFc and rFVIIIFc.</p

Thrombin-mediated release of activated FVIII variants from VWF.

<p>(A) Schematic of the optical biosensor method developed to evaluate the dependence of VWF release rates on thrombin concentration. A detailed description is provided in <i>Materials and Methods</i>. (B–D) Determination of thrombin concentrations corresponding to half-maximal release rates (EC₅₀) for (B) rFVIIIFc, (C) SC rFVIIIFc, and (D) BDD rFVIII. For BDD rFVIII, ordinate values were normalized on a molar basis to account for the molecular weight difference between rFVIIIFc and SC rFVIIIFc (220 kDa), and BDD rFVIII (170 kDa).</p

Non-Reducing SDS-PAGE analysis and schematic of FVIII variants.

<p>(A) SC rFVIIIFc R1645A/R1648A, SC rFVIIIFc, fully processed rFVIIIFc and rFVIIIFc <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0113600#pone.0113600-Peters1" target="_blank">[10]</a> were compared by non-reducing SDS-PAGE. (B) Schematic presentation of SC rFVIIIFc, fully processed rFVIIIFc, and the common thrombin-activated form.</p

SC rFVIIIFc and the rFVIIIFc demonstrated equivalent in vivo efficacy in a HemA mouse bleeding model.

<p>Tail-bleeding of HemA mice was induced via transection of one of the lateral veins at 48 hr post i.v. dosing of SC rFVIIIFc or rFVIIIFc (0.46, 1.38 and 4.6 µg/kg). Mice were monitored for survival and re-bleeding for 24 hr post injury. (A) Schematic experimental design of TVT model. (B) 24 hr post TVT survival curves (C) 24 hr post TVT re-bleeding curves from experimental animals. (D) The linear regression curve of the percentage of protection (survival rate) versus the log (base 10) of the dose was plotted. The ED₅₀ was extrapolated from the curves.</p

Specific activity of SC rFVIIIFc compared to fully processed rFVIIIFc and rFVIIIFc by chromogenic and one-stage (aPTT) assays (n = 3).

<p>Specific activity of SC rFVIIIFc compared to fully processed rFVIIIFc and rFVIIIFc by chromogenic and one-stage (aPTT) assays (n = 3).</p