An affinity gel for the inhibition, binding, and isolation of serine proteases

A preparation procedure for gels for the specific binding and inhibition of serine proteases is described. Phosphoryl trifluoride was synthesized and reacted with two different types of agarose gels, a crosslinked agarose (Sepharose CL-4B) and an agarose containing spacer arms with terminal vicinal-diol groups (a hydrolyzed epoxy-activated Sepharose 6B). The phosphoryl difluoride groups coupled to the gels were, in both cases, further modified by treatment with isopropanol to obtain isopropyl fluorophosphate groups covalently bound to the matrix. It was found that both modified gels absorbed and inhibited plasmin, but that the modified gel with spacer arms was markedly more efficient.