Heating lyophilised factor VIII does not alter its recognition by specific antibodies.
OBJECTIVE
Alterations of factor VIII (FVIII) during preparation procedures can potentially affect its immunogenicity. One method evaluating such alterations could be by determining the reactivity of FVIII with specific antibodies.


METHODS
Since heat treatment is currently used to reduce the risk of viral transmission, we evaluated the immunoreactivity of plasma-derived FVIII before and after heating at different temperatures and for different periods. Freeze-dried FVIII was used for these experiments as part of the validation procedure of a novel FVIII preparation.


RESULTS
Heating FVIII for up to 72 h at 80 degrees C does not alter its reactivity with specific rabbit antibodies or mouse monoclonal antibodies, although some loss of FVIII activity occurred after 72 h. After heating for 2 h at 100 degrees C, a procedure that reduced FVIII activity by about 50%, there were still no significant effects on FVIII reactivity with monoclonal antibodies.


CONCLUSIONS
Freeze-dried preparations of plasma-derived FVIII seem to be resistant to heat-induced structural denaturation.
Gilles
J G
JG
Center for Molecular and Vascular Biology, Katholieke Universiteit Leuven, Belgium.
di Giambattista
M
M
Laub
R
R
Saint-Remy
J M
JM
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions / Q000276:immunology*; D004306:Dose-Response Relationship, Immunologic; D005169:Factor VIII / Q000276:immunology* / Q000302:isolation & purification; D005612:Freeze Drying; D006358:Hot Temperature; D006801:Humans; D051379:Mice; D011817:Rabbits; D013997:Time Factors
D000911:Antibodies, Monoclonal; D005169:Factor VIII