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Thromb Haemost 1979; 42(05): 1388-1397
DOI: 10.1055/s-0038-1657038
Original Article
Schattauer GmbH Stuttgart

A Simple Rapid Method for Isolating Soluble Fibrin Complexes from Fibrinogen by Treatment with Thrombin and t-AMCHA

Shigeru Hayashi
*   The Internal Medicine, Ogikubo Hospital, 3-1-24, Imagawa, Suginami-ku, Tokyo, Japan
,
Kaneo Yamada
**   Department of Pediatrics, St. Marianna Medical University, 2095, Sugao, Takatsuku, Kawasaki-shi, Kanagawa, Japan
› Author Affiliations
Further Information

Publication History

Received 21 July 1978

Accepted 06 June 1979

Publication Date:
23 August 2018 (online)

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Summary

Using treatment with thrombin associated with trans-aminomethylcyclohexane carboxylic acid (t-AMCHA), a simple and rapid method for isolating soluble fibrin complexes (SFC) from fibrinogen in the plasma was developed. By this procedure, the recovery rates of SFC and early FDP (mainly X) increased according to the concentration of t-AMCHA, reaching a maximum at 286 mM t-AMCHA. On the other hand, the recovery rate of fibrinogen remained below 1.5% and that of late FDP was almost 100% at all concentrations of t- AMCHA. These results suggested that SFC and FDP could be isolated from fibrinogen by thrombin and t-AMCHA (286 mM) treatment. Moreover, it was possible to isolate SFC from FDP using gel filtration after treatment with thrombin and t-AMCHA. The SFC could be quantified by assay of the eluted fractions containing SFC by the staphylococcal clumping test.

 

  • References

  • 1 Fletcher AP, Alkjaersig NK. 1971; Blood hypercoagulability, intravascular coagulation and thrombosis. New diagnostic concepts Thromb. Diath. Haemorrh. 45 (Suppl.) 389
    PubMedGoogle Scholar
  • 2 Hawiger J, Niewiarowski S, Gurewich V, Thomas DP. 1970; Measurement of fibrinogen and fibrin degradation products in serum by staphylococcal clumping test. J. Lab. Clin. Med. 75: 993
    PubMedGoogle Scholar
  • 3 Hayashi S. 1979; Studies on the significance of serum soluble fibrin in nephritis, with special reference to the technique of staphylococcal clumping. Acta Haem. Jap. 42: 53
    PubMedGoogle Scholar
  • 4 Heene DL, Matthias FR. 1973; Adsorption of fibrinogen derivatives on insolubilized fibrinogen and fibrin monomer. Thrombos. Res. 2: 137
    CrossrefPubMedGoogle Scholar
  • 5 Von Hugo R, Hafter R, Stemberger A, Graefe H. 1975; Soluble fibrin monomer complexes demonstrated by agarose gel filtration and by adsorption on insolubilized fibrinogen. Thromb. Diath. Haemorrh. 34: 216
    PubMedGoogle Scholar
  • 6 Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. 1951; Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193: 265
    PubMedGoogle Scholar
  • 7 Matthias FR, Reinicke R, Heene DL. 1977; Affinity chromatography and quantification of soluble fibrin from plasma. Thrombos. Res. 10: 365
    CrossrefPubMedGoogle Scholar
  • 8 Weber K, Osborn M. 1969; The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J. Biol. Chem. 244: 4406
    PubMedGoogle Scholar