PEPTIDE AFFINITY-CHROMATOGRAPHY OF HUMAN CLOTTING FACTOR-VIII SCREENING OF THE VWF-BINDING DOMAIN

The region of von Willebrand factor, which is involved in the complex formation with factor VIII, was used to generate a panel of octapeptid es. A peptide ladder was generated from the von Willebrand factor regi on aa40 to aa100 and was synthesized on cellulose membranes by spot te chnology. Four peptides with affinity for factor VIII were identified by incubation with plasma derived factor Vm and recombinant factor VII I. The peptides denoted as 010 (LCPPGMVRHE), 011 (RCPCFHQGK), 014 (CFH QGKEYA) and 015 (RDRKWNCTDHVC) were further characterized by real-time interaction analysis and small scale affinity chromatography. Biotiny lated peptides were used for blotting assays. These experiments showed that the peptides are directed against the light chain of FVIII. We c onsider these peptides as valuable tools for in situ labeling and also as ligands suitable for affinity chromatography. (C) 1998 Elsevier Sc ience B.V. All rights reserved.