The present invention relates to the field of therapeutic proteins, in particular, to recombinant coagulation factors. It provides a
recombinant Factor VIII (FVIII)
protein comprising specific point mutations at defined positions, which serve to reduce the
immunogenicity of said FVIII
protein, wherein the Factor VIII
protein substantially retains its coagulant activity. The invention further provides nucleic acids encoding said de-immunized protein,
cell lines and methods of recombinant preparation as well as pharmaceutical compositions comprising the recombinant FVIII of the invention, whichare advantageous for use in treatment of patients with Hemophilia A, particularly those who have not yet been treated with a FVIII product. Additionally, it can be a safe alternative for
previously treated patients and even for patients who have developed an immune-response to FVIII, e.g., for immune-tolerance-
induction therapy (ITI / ITT) or rescue ITI. The invention also provides an
assay for determining
immunogenicity of a protein.