PROCESSING OF FUSION PROTEINS WITH IMMOBILIZED FACTOR XA

Factor Xa, with a cellulose-binding domain (CBD) fused to the C-terminus of the heavy chain (FXa-CBD), is active in solution and when immobilized on cellulose. A second derivative of factor Xa in which a hexahistidine tail is fused to the C-terminus of the. heavy chain (FXa-H-6) also retains activity when immobilized, in this case on Ni2+-NTA agarose. The stabilities and activities of FXa-CBD and FXa-H-6 immobilized on cellulose and Ni2+-NTA agarose, respectively, are similar. Immobilized factor Xa derivatives can be used to remove affinity tags from appropriate fusion proteins without contaminating the desired product with factor Xa.