Effects of genetic fusion of factor IX to albumin on in vivo clearance in mice and rabbits.
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 (t(0.5)) did not differ amongst the three proteins. Two minutes after injection, >40% of the injected MFUST was found in the liver, compared with <10% of either MSAT or pd-MSA. In rabbits, the AUC for MFUST was reduced compared to MIXT, MSAT, or pd-MSA, while the t(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.
Sheffield
William P
WP
Department of Pathology and Molecular Medicine, McMaster University, Hamilton, Ontario, Canada. sheffiel@mcmaster.ca
Mamdani
Asif
A
Hortelano
Gonzalo
G
Gataiance
Sharon
S
Eltringham-Smith
Louise
L
Begbie
Megan E
ME
Leyva
Rina A
RA
Liaw
Peter S
PS
Ofosu
Frederick A
FA
D000818:Animals; D001777:Blood Coagulation; D005164:Factor IX / Q000493:pharmacokinetics*; D006207:Half-Life; D051379:Mice; D018345:Mice, Knockout; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000493:pharmacokinetics*; D012709:Serum Albumin / Q000493:pharmacokinetics*; D014018:Tissue Distribution
D011993:Recombinant Fusion Proteins; D012709:Serum Albumin; D005164:Factor IX