Properties of a Highly Purified Human Plasma Factor IX:c
Therapeutic Concentrate Prepared by Conventional Chromatography

Burnouf, T.; Michalski, C.; Goudemand, M.; Huart, J.J.

doi:10.1159/000461052

Cited by 32 publications

(55 citation statements)

References 11 publications

(31 reference statements)

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“…The FX-containing fraction was obtained during the isolation of factor IX (FIX) and corresponded to the 0.28 mmol/L NaCl eluate, 17 which was applied onto a ceramic hydroxyapatite column (Bio-Rad) equilibrated with 10 mmol/L K 2 HPO 4 and 75 mmol/L NaCl (pH 8). After a wash with 30 mmol/L K 2 HPO 4 and 0.25 mmol/L NaCl (pH 8), FX was eluted with 0.5 mmol/L K 2 HPO 4 (pH 8) and then dialyzed overnight against 20 mmol/L Tris and 0.15 mmol/L NaCl (pH 7) at 4°C.…”

Section: Human Fx Isolationmentioning

confidence: 99%

Vascular Endothelial Growth Factor Production by Fibroblasts in Response to Factor VIIa Binding to Tissue Factor Involves Thrombin and Factor Xa

Ollivier

Chabbat

Herbert

et al. 2000

ATVB

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Tissue factor (TF) assembled with activated factor VII (FVIIa) initiates the coagulation cascade. We recently showed that TF was essential for FVIIa-induced vascular endothelial growth factor (VEGF) production by human fibroblasts. We investigated whether this production resulted from TF activation by its binding to FVIIa or from the production of clotting factors activated downstream. Incubation of fibroblasts with a plasma-derived FVIIa concentrate induced the generation of activated factor X (FXa) and thrombin and the secretion of VEGF, which was inhibited by hirudin and FXa inhibitors. By contrast, the addition of recombinant FVIIa to fibroblasts did not induce VEGF secretion unless factor X was present. Moreover, thrombin and FXa induced VEGF secretion and VEGF mRNA accumulation, which were blocked by hirudin and FXa inhibitors, respectively. The effect of thrombin was mediated by its specific receptor, protease-activated receptor-1; in contrast, the effect of FXa did not appear to involve effector cell protease receptor-1, because it was not affected by an anti-effector cell protease receptor-1 antibody. An increase in intracellular calcium with the calcium ionophore A23187 or intracellular calcium chelation by BAPTA-AM had no effect on either basal or FXa-induced VEGF secretion, suggesting that the calcium signaling pathway was not sufficient to induce VEGF secretion. Finally, FVIIa, by itself, had no effect on mitogen-activated protein (MAP) kinase activation, contrary to thrombin and FXa, which activate the p44/p42 MAP kinase pathway, as shown by the blocking effect of PD 98059 and by Western blotting of activated MAP kinases. These findings indicate that FVIIa protease induction of VEGF expression is mediated by thrombin and FXa generated in response to FVIIa binding to TF-expressing fibroblasts; they also exclude a direct signaling involving MAP kinase activation via the intracellular domain of TF when expressed by these cells. (Arterioscler Thromb Vasc Biol. 2000;20:1374-1381.) Key Words: vascular endothelial growth factor tissue factor activated factor VII fibroblasts proteases T issue factor (TF), the cell-surface receptor for activated factor VII (FVIIa), is the primary regulator of blood coagulation. The TF-FVIIa complex cleaves and activates factors IX and X into factors IXa and factor Xa (FXa), respectively, which lead to thrombin generation. 1 Beyond its role as a procoagulant activator, TF participates in other cellular processes, including metastasis, 2 tumor-associated angiogenesis, 3 and embryogenesis. 4 Accordingly, several cell functions are modified in response to FVIIa binding to TF, its receptor. These include intracellular signaling, 5 activation of the p44/42 mitogen-activated protein (MAP) kinase pathway , 6 induction of tyrosine phosphorylation in monocytes, 7 upregulation of poly(A) polymerase, 8 cell spreading and phosphorylation of focal adhesion kinase, 9 enhanced expression of urokinase receptor by pancreatic cancer cell lines, 10 and vascular endothelial growth factor (VEGF)...

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Section: Human Fx Isolationmentioning

confidence: 99%

Vascular Endothelial Growth Factor Production by Fibroblasts in Response to Factor VIIa Binding to Tissue Factor Involves Thrombin and Factor Xa

Ollivier

Chabbat

Herbert

et al. 2000

ATVB

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“…The method includes a sol vent/detergent step for the inactivation step of lipid-enveloped viruses [19] . The purified product has a typical specific activity of 110 IU/mg of protein and was formulated with 0.1% sucrose (= 0.03 M) in addi tion to the constituents described by Burnouf et al [17], High-purity factor IX was prepared from a factor II, IX and X con centrate by ion-exchange chromatography with DEAE-Sepharose FF [20] , followed by affinity chromatography using heparin-Sepharose [21] . Solvent-detergent treatment was included between these steps to inactivate enveloped viral contaminants.…”

Section: Concentrate Samples and Process Modelsmentioning

confidence: 99%

Effect of Terminal (Dry) Heat Treatment on Non-Enveloped Viruses in Coagulation Factor Concentrates

Hart¹,

Hart²,

Crossley³

et al. 1994

Vox Sanguinis

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Terminal dry heat treatment effectively inactivated hepatitis A virus (HAV) and canine parvovirus added to high-purity factor VIII. After 24 h at 80°C, HAV in- fectivity was reduced by ≥4.3 log(10) TCID(50), as measured in a newly developed infectivity assay. The same reduction in virus titer was achieved after 2 h and before 6 h at 90°C. Inactivation of hepatitis A virus was also seen in the freezedrying step prior to heat treatment with an approximately 2.0 log(10) reduction in titer. Similar results were obtained with a high-purity factor IX concentrate. Canine parvovirus was also inactivated at both temperatures, with residual infectivity being undetected after 48 hat 80°C or 10 hat 90°C. Canine parvovirus was not affected by lyophilisation. Canine parvovirus measurements by PCR did not reflect the levels of infectivity measured by the tissue-culture-based method. The addition of the terminal dry heat treatment to solvent/detergent could effectively eliminate the potential contamination of solvent/detergent-treated coagulation factor concentrates by non-lipid-enveloped vimses. However, careful evaluation for any increased induction of non-antigens for factor VIII, as a consequence of such treatment, is needed before use in patients can be recommended.

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“…Three-factor PCC are now gradually being replaced by high-purity FIX concentrates, which are obviously less thrombogenic [15][16][17][18][19].…”

Section: Introductionmentioning

confidence: 99%

Factor VII and Activated- Factor-VII Content of Prothrombin Complex Concentrates

Hellstern¹,

Beeck²,

Fellhauer³

et al. 1997

Vox Sanguinis

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Background and objectives: The aim of this study was to determine the potencies of factor VII (FVII) and of activated FVII (FVIIa) in prothrombin complex concentrates (PCC). Materials and methods: We examined 56 lots of PCC from 5 manufacturers. Three brands were licensed preparations, and 1 product scries had been involved in thromboembolic complications. FVII and FVIIa were measured using a two-stage amidolytic assay and a specific clotting assay, respectively. We also quantified FVII clotting activity by a one-stage assay reflecting a mixture of FVII zymogen and FVIIa. Results: All PCC contained substantial amounts of FVII, and FVIIa could be detected in all lots. There were marked differences between manufacturers and some significant variabilities between batches. The two lots involved in thromboembolic events contained considerably more FVIIa than the PCC still licensed. The lowest FVIIa potencies were observed in an experimental product series, indicating that PCC can be produced without activation of FVII during the manufacturing process. Conclusion: FVIIa is present in all PCC containing FVII. High FVIIa potencies may contribute to the thrombogenic potential of these preparations, and determination of FVIIa potencies should be included in the in vitro characterization of PCC.

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Properties of a Highly Purified Human Plasma Factor IX:c Therapeutic Concentrate Prepared by Conventional Chromatography

Cited by 32 publications

References 11 publications

Vascular Endothelial Growth Factor Production by Fibroblasts in Response to Factor VIIa Binding to Tissue Factor Involves Thrombin and Factor Xa

Vascular Endothelial Growth Factor Production by Fibroblasts in Response to Factor VIIa Binding to Tissue Factor Involves Thrombin and Factor Xa

Effect of Terminal (Dry) Heat Treatment on Non-Enveloped Viruses in Coagulation Factor Concentrates

Factor VII and Activated- Factor-VII Content of Prothrombin Complex Concentrates

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