Chromatography of the VIII/vWF complex on dextran sulphate sepharose

The applicability of Dextran Sulphate (DS) agarose in the purification of the VIII/vWf complex from cryoprecipitate was investigated. The elution profiles indicate high affinity of the VIII/vWf complex for the matrix. With application of combined salt/pH gradients good resolution of VIII/vWf from most of the applied proteins can be achieved. VIII/vWf normally elutes as a single component (70-90% vWf:Ag and vWf:RCo) with modest yields of factor VIII (30-40%) VIII:C and 50-70% VIII:Ag) providing calcium ions are present. Addition of L-lysine, Histidine and Arginine (all 1.0M) inhibit binding of proteins to the matrix. Alternatively, higher yields of VIII:C can be obtained (80% VIII:C and 100% VIII:Ag) by the application of L-lysine gradients (0-0.6M) but with loss of resolution from Fn and slight dissociation of VIII:C from vWf:Ag.