Abstract | INTRODUCTION: MATERIALS AND METHODS: A novel method using magnetic chromatography was used to isolate catabolic organelle (CO) fractions from mouse liver following injection of superparamagnetic dextran (SPD)-coated iron oxide particles and rFVIIa. The effect of co-circulating SPD particles on rFVIIa pharmacokinetic (PK) parameters was evaluated by ELISA. Cryo-immuno transmission electron microscopy (TEM) was used to study hepatic distribution of SPD particles and rFVIIa. The isolated hepatic CO fractions were characterized using Western Blotting (WB). RESULTS: Cryo-immuno TEM of the liver confirmed hepatic co-localisation of SPD particles and rFVIIa in identical endosomes and lysosomes of both hepatocytes and Kupffer cells. SPD particles did not affect the PK parameters of rFVIIa. WB analysis of plasma and CO fractions detected rFVIIa as the full-length protein and also in high molecular weight (HMW) complexes with ATIII and α-2 macroglobulin (α-2M). CONCLUSIONS: Following injection, both hepatocytes and Kupffer cells appeared to be involved in the hepatic clearance and metabolism of both full-length rFVIIa and rFVIIa in complex with at least two plasma protease inhibitors; ATIII and α-2M.
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Authors | T Seested, R S Appa, E I Christensen, Y A Ioannou, T N Krogh, D M Karpf, H M Nielsen |
Journal | Thrombosis research
(Thromb Res)
Vol. 127
Issue 4
Pg. 356-62
(Apr 2011)
ISSN: 1879-2472 [Electronic] United States |
PMID | 21262526
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Copyright | Copyright © 2010 Elsevier Ltd. All rights reserved. |
Chemical References |
- Dextrans
- Ferric Compounds
- Protease Inhibitors
- Recombinant Proteins
- ferric oxide
- recombinant FVIIa
- Factor VIIa
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Topics |
- Animals
- Chromatography
(methods)
- Dextrans
(chemistry)
- Factor VIIa
(administration & dosage, metabolism, pharmacokinetics)
- Ferric Compounds
(chemistry)
- Hepatocytes
(metabolism)
- Injections, Intravenous
- Liver
(metabolism)
- Magnetics
- Male
- Mice
- Protease Inhibitors
(metabolism)
- Recombinant Proteins
(administration & dosage, metabolism, pharmacokinetics)
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