It is found that clotting of fibrinogen by thrombin does not occur on the acid side of the isoelectric point of the fibrinogen. At such pH values, however, a primary reaction between thrombin and fibrinogen takes place, leading to the formation of profibrin, a compound of thrombin and fibrinogen.
At pH values at which clotting is possible, fibrinogen is negatively, thrombin positively charged, whereas profibrin has a pattern of positive and negative charges.
The primary reaction, the formation of profibrin by combination of thrombin and fibrinogen, is inhibited by urea but not by neutral salts. The combination of thrombin with fibrinogen most probably takes place by hydrogen bonds.
The second reaction, the polymerisation of profibrin to fibrin, is inhibited by neutral salts in the same way as complex or autocomplex coacervates. It is caused therefore by electrostatic attraction between the positive and the negative charges of the profibrin.