Generation of Thrombin from Blood for Therapeutic Use

Sternberger (1) described a method for the preparation of human thrombin in the first volume of the Journal of Clinical Pathology. This study described the preparation of thrombin from human blood in relatively small quantities and could be used for the preparation of autologous thrombin.   Thrombin is generated from oxalated whole blood in the presence of calcium ions, ethanol, and human milk; thromboplastic activity in provided from whole milk (human milk said to have a higher level of thromboplastic activity than bovine milk).   The thrombin is precipitated with acacia and is stable for an extended period of time.  The material, a paste of thrombin and acacia, was shown to be clinically useful.  This paper has not been extensively cited; Godal and coworkers (2) raised issues with Sternberger's later concept of continuous thrombin formation but did report using thrombin prepared by the Sternberger method.  Acacia does accelerate fibrinogen clotting (3) by accelerating polymerization in a manner similar to that seen for other colloids such as poly(ethylene)glycol (4).   Boyles and coworkers presented evidence  to support the concept that the formation of a fibrin clot was the result of the conversion of fibrinogen to fibrin in an enzyme-catalyzed reaction and not a result of fibrinogen denaturation.  The formation of fibrin does have characteristics of protein denaturation and thrombin was described as a denaturase in early studies.   Erwin Chargaff and Aaron Bendich (5) described the formation of coagulated fibrinogen, as differentiated from fibrin formed by thrombin, by a variety of organic compounds including ninhydrin and chloramine-T.   These investigators also mentioned the description of thrombin as a denaturase (6).  As a historical note, both Chargaff and Bendich went on to distinguished careers at Columbia University in work totally unrelated to fibrinogen.  Chargaff played an important role in the description of deoxyribonucleic acid as described in Jim Watson's book, The Double Helix.

References
1.  Sternberger,  L.A.,  Haemostasis with an easily prepared stable thrombin solution, J.Clin.Pathol. 1, 229-231, 1948.
2.  Godal, H.C., Hasselback, R., and Hjort, P.F., The recovery of thrombin from plasma, Scand.J.Clin.Lab.Invest. 13, 532-533, 1961.
3.  Boyles, P.W., Ferguson, J.H., and Muehlke, P.H., Mechanisms involved in fibrin formation, J.Gen.Physiol. 34, 493-513, 1951.
4.  Fenton, J.W., 2nd and Fasco, M.J., Polyethylene glycol 6,000 enhancement of the clotting of fibrinogen solutions in visual and mechanical assays, Thromb.Res. 4, 809-817, 1974.
5. Chargaff, E. and Bendich, A., On the coagulation of fibrinogen, J.Biol.Chem. 149, 93-110, 1943.
6. Wohlisch, E. and Juhling, L., Thrombin as denaturase and its relation to papain, Biochemische Zeitschrift 297, 353-368, 1938.