Large-scale preparation of human thrombin: polyethylene glycol potentiates the factor Xa-mediated activation of prothrombin.
We investigated the ability of polyethylene glycol 4000 to accelerate thrombin generation in a mixture of prothrombin and factor X at concentrations of 1-30%. In the presence of 5 mM of CaCl2, polyethylene glycol 4000 promoted prothrombin activation at concentrations above 1%. The peak of activation was seen at levels of 14 and 20% of polyethylene glycol 4000. The effect of the polyethylene glycol was remarkably dependent on its molecular weight; molecular weights greater than 2000 were required for accelerating thrombin generation. Under optimal conditions, polyethylene glycol 4000, in the presence of CaCl2, promoted conversion of all of the prothrombin into thrombin and its derivatives. We conclude that polyethylene glycol 4000, at concentrations ranging from 14 to 20%, effectively accelerates thrombin generation in the presence of 5 mM of CaCl2. This new method for preparing thrombin is based on the use of polyethylene glycol 4000 and CaCl2 and is applicable to the manufacture of thrombin.
Kaetsu
H
H
The Chemo-Sero-Therapeutic Research Institute, Kumamoto, Okubo, Japan. QYN00365@niftyserve.or.jp
Mizuguchi
J
J
Hamamoto
T
T
Kamimura
K
K
Yoshida
Y
Y
Nakagaki
T
T
Ogata
Y
Y
Miyamoto
S
S
Funatsu
A
A
D002122:Calcium Chloride / Q000032:analysis; D004345:Drug Industry; D004789:Enzyme Activation / Q000187:drug effects; D015951:Factor Xa / Q000187:drug effects / Q000378:metabolism; D005990:Glycerol / Q000494:pharmacology; D006801:Humans; D008420:Manufactured Materials; D008970:Molecular Weight; D009994:Osmolar Concentration; D011092:Polyethylene Glycols / Q000032:analysis / Q000494:pharmacology; D011516:Prothrombin / Q000187:drug effects / Q000378:metabolism; D013696:Temperature; D013917:Thrombin / Q000187:drug effects / Q000378:metabolism*; D013997:Time Factors
D011092:Polyethylene Glycols; D011516:Prothrombin; D013917:Thrombin; D015951:Factor Xa; D002122:Calcium Chloride; D005990:Glycerol