title: Isolation of plasma kallikrein by high efficiency affinity chromatography and its characterization.
abstract: We established a simple method for the purification of human plasma kallikrein (PK) by affinity chromatography and characterized it by analytical reverse phase-HPLC and Time of Flight Mass Spectroscopy (TOF-MS). The affinity resin (PKSI-Toyopearl) was synthesized using a selective synthetic inhibitor of plasma kallikrein (PKSI-527) as a ligand. The resin was found to have the highest efficiency in PK purification when the coupling ratio of PKSI-527 per resin was 9-14 micromol/g. PK was purified 466-fold with a yield of 83% from acetone-activated human plasma by affinity chromatography. The purity of PK thus obtained was confirmed by reverse phase-HPLC with a linear gradient of acetonitrile. The molecular weight of the purified PK was determined to be 86,151 by TOF-MS.
authors.affiliation: Faculty of Pharmaceutical Sciences, Kobe Gakuin University, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Tada
authors.forename: Y
authors.initials: Y
authors.lastname: Tsuda
authors.forename: K
authors.initials: K
authors.lastname: Wanaka
authors.forename: A
authors.initials: A
authors.lastname: Hijikata-Okunomiya
authors.forename: N
authors.initials: N
authors.lastname: Horie
authors.forename: U
authors.initials: U
authors.lastname: Okamoto
authors.forename: S
authors.initials: S
authors.lastname: Okamoto
authors.forename: Y
authors.initials: Y
authors.lastname: Okada
mesh_terms: D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007610:Kallikreins / Q000032:analysis / Q000097:blood / Q000302:isolation & purification*; D013058:Mass Spectrometry; D008970:Molecular Weight
chemical_list: D007610:Kallikreins