PURIFICATION METHOD FOR HUMAN PLASMA KALLIKREIN BY A NEW AFFINITY-CHROMATOGRAPHY

We synthesized a new affinity gel (PKSI-Toyopearl) using a selective s ynthetic inhibitor of plasma kallikrein (PKSI-527) as an affinity liga nd, and employed it for the rapid purification of plasma kallikrein fr om human plasma. Human plasma activated with kaolin after acid treatme nt was applied to a PKSI-Toyopearl column. Adsorbed protein was eluted with 50 mM glycine-hydrochloric acid buffer (pH 3.0). Plasma kallikre in was purified 181-fold with a yield of 85% from the kaolin-activated plasma. Further purification was performed by chromatography on a DEA E-Toyopearl 650M column. Plasma kallikrein,vas finally purified 1720-f old with a 63% yield by these procedures. On sodium dodecyl sulfate (S DS) polyacrylamide gel electrophoresis, a band was observed at approxi mately 88 kDa. These findings indicate that PKSI-Toyopearl is a valuab le tool for the purification of plasma kallikrein from human plasma.