Purification method for human plasma kallikrein by a new affinity chromatography.
We synthesized a new affinity gel (PKSI-Toyopearl) using a selective synthetic inhibitor of plasma kallikrein (PKSI-527) as an affinity ligand, and employed it for the rapid purification of plasma kallikrein from human plasma. Human plasma activated with kaolin after acid treatment was applied to a PKSI-Toyopearl column. Adsorbed protein was eluted with 50 mM glycine-hydrochloric acid buffer (pH 3.0). Plasma kallikrein was purified 181-fold with a yield of 85% from the kaolin-activated plasma. Further purification was performed by chromatography on a DEAE-Toyopearl 650M column. Plasma kallikrein was finally purified 1720-fold with a 63% yield by these procedures. On sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis, a band was observed at approximately 88 kDa. These findings indicate that PKSI-Toyopearl is a valuable tool for the purification of plasma kallikrein from human plasma.
Tada
M
M
Kobe Research Projects on Thrombosis and Haemostasis, Japan.
Wanaka
K
K
Okamoto
S
S
Okamoto
U
U
Nakaya
Y
Y
Horie
N
N
Hijikata-Okunomiya
A
A
Tsuda
Y
Y
Okada
Y
Y
D001769:Blood; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007610:Kallikreins / Q000302:isolation & purification* / Q000378:metabolism; D013379:Substrate Specificity
D007610:Kallikreins