Argatroban-coupled Affi-Gel matrix for the purification of thrombin from plasma.
Sometimes it is necessary to obtain thrombin from limited amounts of human plasma for laboratory assay. None of the available purification methods easily deals with this subject. The procedure described in the present paper uses a readily available pharmaceutical agent, argatroban, to construct an affinity matrix. Argatroban has a high affinity for thrombin and its thrombin binding is reversible. Prothrombin derived from a Ba(2+) precipitate of human plasma is used as the starting material. The crude prothrombin can be bulk activated to thrombin using taipan-snake (Oxyuranus scutellatus) venom and bound to the argatroban-coupled matrix without further processing steps. The thrombin product eluted from the argatroban matrix is very pure as judged by high specific activity and by electrophoresis. This purification scheme is rapid, yielding purified thrombin within 2 days.
Lefkowitz
Jerry B
JB
Department of Pathology, University of Colorado Health Sciences Center, RC1, P18-5124, Aurora, CO 80045-0508, U.S.A. Jerry.Lefkowitz@UCHSC.edu
D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives; D017815:Elapidae; D004591:Electrophoresis, Polyacrylamide Gel; D005782:Gels; D010875:Pipecolic Acids / Q000737:chemistry*; D010949:Plasma / Q000378:metabolism*; D013449:Sulfonamides; D013917:Thrombin / Q000302:isolation & purification* / Q000378:metabolism
D005782:Gels; D010875:Pipecolic Acids; D013449:Sulfonamides; D001120:Arginine; D013917:Thrombin; C031942:argatroban