Increase in fibrinogen and fibrin-related antigen in human serum due to in vitro lysis of fibrin by thrombin.
In vitro lysis of fibrin, as indicated by increased fibrinogen-fibrin-related antigen (FR-antigen) in serum is usually seen when whole blood, or plasma, or highly purified fibrinogen prepared by several different procedures is clotted and kept at temperatures above 0 degrees C. This increase is both time and temperature dependent, occurs despite the addition of various plasmin and cathepsin inhibitors, and is probably caused by thrombin evolved during clotting and/or added in vitro. In these experiments, the FR-antigen was measured by a sensitive, reproducible hemagglutination inhibition immunoassay adapted to the AutoAnalyzer. Serum from whole blood contained more than serum from plasma, and fibrin rather than fibrinogen proved to be essential for the in vitro lysis. The phenomenon was also caused by Arvin or Reptilase, suggesting that splitting of one or more arginine or lysine bonds in fibrin may be at least partially responsible. To obtain minimal levels of FR-antigen (< 0.5 mug/ml), plasma is clotted for 4 hr at 0 degrees C with 1.0-5.0 U/ml thrombin, CaCl(2) (0.0125 mole/liter), and epsilon aminocaproic acid (0.05 mole/liter). Slightly higher levels, probably adequate for clinical diagnosis, are obtained by 10-30 min clotting at room temperature. Since endogenous and/or exogenous thrombin is essential for the collection of serum FR-antigen, all the FR-antigen found in normal serum probably results from an irreducible amount of in vitro lysis rather than from continuous intravascular clotting and fibrinolysis.
D000614:Aminocaproates; D000941:Antigens / Q000032:analysis*; D001322:Autoanalysis; D001565:Benzoates; D002118:Calcium; D002403:Cathepsins / Q000037:antagonists & inhibitors; D005337:Fibrin / Q000032:analysis*; D005340:Fibrinogen / Q000032:analysis* / Q000302:isolation & purification; D005342:Fibrinolysis; D006146:Guanidines; D006385:Hemagglutination Inhibition Tests; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay; D007120:Immunochemistry; D066298:In Vitro Techniques; D009574:Nitro Compounds; D009994:Osmolar Concentration; D010958:Plasminogen; D013917:Thrombin; D014105:Tosyl Compounds
D000614:Aminocaproates; D000941:Antigens; D001565:Benzoates; D006146:Guanidines; D009574:Nitro Compounds; D014105:Tosyl Compounds; D005337:Fibrin; D005340:Fibrinogen; D010958:Plasminogen; D002403:Cathepsins; D013917:Thrombin; D002118:Calcium