Effects of genetic fusion of factor IX to albumin on in vivo clearance in mice and rabbits

William P. Sheffield, Asif Mamdani, Gonzalo Hortelano, Sharon Gataiance, Louise Eltringham-Smith, Megan E. Begbie, Rina A. Leyva, Peter S. Liaw, Frederick A. Ofosu

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Individuals with haemophilia B require replacement therapy with recombinant or plasma-derived coagulation factor IX (fIX). More benefit per injected dose might be obtained if fIX clearance could be slowed. The contribution of overall size to fIX clearance was explored, using genetic fusion to albumin. Recombinant murine fIX (MIX), and three proteins with C-terminal epitope tags were expressed in HEK 293 cells: tagged MIX (MIXT), tagged mouse serum albumin (MSAT) and MFUST, in which MIX and MSAT were fused in a single polypeptide chain. Proteins MFUST and MIXT were two- to threefold less active in clotting assays than MIX. In mice, the area under the clearance curve (AUC) was reduced for MFUST compared with MSAT or plasma-derived MSA (pd-MSA); the terminal catabolic half-life (t0.5) did not differ amongst the three proteins. Two minutes after injection, >40% of the injected MFUST was found in the liver, compared with 0.5 of the fusion protein fell between that of MIXT and MSAT or pd-MSA. Similar results were obtained with non-radioactive fused or non-fused recombinant human fIX in fIX knockout mice. The clearance behaviour of the fusion protein thus more closely resembled that of fIX than that of albumin despite a modest increase in terminal half-life, suggesting that fIX-specific interactions that are important in determining clearance were maintained in spite of the increased size of the fusion protein.

Original languageEnglish
Pages (from-to)565-573
Number of pages9
JournalBritish Journal of Haematology
Volume126
Issue number4
DOIs
Publication statusPublished - Aug 2004
Externally publishedYes

Fingerprint

Factor IX
Albumins
Rabbits
Proteins
Half-Life
Hemophilia B
HEK293 Cells
Protein C
Knockout Mice
Serum Albumin
Area Under Curve
Epitopes
Peptides
Injections
Liver

Keywords

  • Albumin
  • Clearance
  • Factor IX
  • Fusion protein

ASJC Scopus subject areas

  • Hematology

Cite this

Sheffield, W. P., Mamdani, A., Hortelano, G., Gataiance, S., Eltringham-Smith, L., Begbie, M. E., ... Ofosu, F. A. (2004). Effects of genetic fusion of factor IX to albumin on in vivo clearance in mice and rabbits. British Journal of Haematology, 126(4), 565-573. https://doi.org/10.1111/j.1365-2141.2004.05106.x

Effects of genetic fusion of factor IX to albumin on in vivo clearance in mice and rabbits. / Sheffield, William P.; Mamdani, Asif; Hortelano, Gonzalo; Gataiance, Sharon; Eltringham-Smith, Louise; Begbie, Megan E.; Leyva, Rina A.; Liaw, Peter S.; Ofosu, Frederick A.

In: British Journal of Haematology, Vol. 126, No. 4, 08.2004, p. 565-573.

Research output: Contribution to journalArticle

Sheffield, WP, Mamdani, A, Hortelano, G, Gataiance, S, Eltringham-Smith, L, Begbie, ME, Leyva, RA, Liaw, PS & Ofosu, FA 2004, 'Effects of genetic fusion of factor IX to albumin on in vivo clearance in mice and rabbits', British Journal of Haematology, vol. 126, no. 4, pp. 565-573. https://doi.org/10.1111/j.1365-2141.2004.05106.x
Sheffield, William P. ; Mamdani, Asif ; Hortelano, Gonzalo ; Gataiance, Sharon ; Eltringham-Smith, Louise ; Begbie, Megan E. ; Leyva, Rina A. ; Liaw, Peter S. ; Ofosu, Frederick A. / Effects of genetic fusion of factor IX to albumin on in vivo clearance in mice and rabbits. In: British Journal of Haematology. 2004 ; Vol. 126, No. 4. pp. 565-573.
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