Intrinsic stability and functional properties of disulfide bond-stabilized coagulation factor VIIIa variants

A. Gale (1) , K.-P. Radtke (2) , M. Cunningham (2) , D. Chamberlain (2) , Jean-Luc Pellequer (3, 4) , J. Griffin (1)

1 Division of Biochemistry, Department of Molecular and Experimental Medicine
2 Biological Products Division
3 SBTN - Service de Biochimie et Toxicologie Nucléaire
4 IBS - UMR 5075 - Institut de biologie structurale

The utility of purified coagulation factor (F)VIII for treatment of hemophilia A is limited in part by its instability following activation by thrombin, which is caused by spontaneous dissociation of the A2 domain from the activated FVIII (FVIIIa) heterotrimer. To prevent this A2 domain dissociation in FVIIIa, we previously engineered a cysteine pair (C664-C1826) in recombinant FVIII that formed a disulfide bond cross-linking the A2 domain in the heavy chain to the A3 domain in the light chain. This engineered disulfide bond resulted in a more stable FVIIIa.

Here, we characterize the functional parameters of C664-C1828 FVIII and of a new disulfide bond-stabilized FVIII (C662-C1828 FVIII).

Domaines: Biologie structurale [q-bio.BM]

A. Gale, K.-P. Radtke, M. Cunningham, D. Chamberlain, Jean-Luc Pellequer, et al.. Intrinsic stability and functional properties of disulfide bond-stabilized coagulation factor VIIIa variants. Journal of Thrombosis and Haemostasis, 2006, 4 (6), pp.1315-1322. ⟨10.1111/j.1538-7836.2006.01951.x⟩. ⟨hal-03242813⟩