Factor VIII performs a critical role in the blood coagulation cascade but is extremely labile. We have carried out chemical studies on a fully functional recombinant Factor VIII (rFVIII) using a variety of protein modifying agents, some of which were bifunctional. Thiol-specific maleimides cause no activity loss despite a substantial decrease in rFVIII's free thiol content. Mild oxidation procedures had either neutral or adverse effects on activity. Amino-specific reagents led to significant losses of rFVIII procoagulant activity. It appears that free amino groups are essential for Factor VIII's function while some at least of its many free thiol groups are not. None of these derivatives was any less labile than unmodified rFVIII. Systematic chemical modification of important proteins in this way can give useful insights into appropriate protein engineering strategies.