Abstract
Circulating soluble fibrin, observed in the blood of patients with ongoing intravascular coagulation, is generated from the plasma protein fibrinogen by the limited proteolytic action of thrombin. We report the production of a monoclonal antibody that discriminates between fibrin and fibrinogen in blood. The synthetic hexapeptide Gly-Pro-Arg-Val-Val-Glu, representing the amino terminus of the alpha chain of human fibrin, was used as immunogen. This hexapeptide is located within the A alpha chain of fibrinogen but becomes the amino terminus of the fibrin alpha chain, after fibrinopeptide A is removed by the action of thrombin, and thus becomes accessible for antibody binding. The monoclonal antibody we have prepared can discriminate between fibrin and fibrinogen and thus can be used in assay systems to quantitate soluble fibrin or, potentially, to image fibrin-rich thrombi.
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Selected References
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- BETTELHEIM F. R. The clotting of fibrinogen. II. Fractionation of peptide material liberated. Biochim Biophys Acta. 1956 Jan;19(1):121–130. doi: 10.1016/0006-3002(56)90393-6. [DOI] [PubMed] [Google Scholar]
- Blombäck B., Hessel B., Hogg D., Therkildsen L. A two-step fibrinogen--fibrin transition in blood coagulation. Nature. 1978 Oct 12;275(5680):501–505. doi: 10.1038/275501a0. [DOI] [PubMed] [Google Scholar]
- FERRY J. D. Polymerization of fibrinogen. Physiol Rev. 1954 Oct;34(4):753–760. doi: 10.1152/physrev.1954.34.4.753. [DOI] [PubMed] [Google Scholar]
- Fletcher A. P., Alkjaersig N., O'Brien J., Tulevski V. G. Blood hypercoagulability and thrombosis. Trans Assoc Am Physicians. 1970;83:159–167. [PubMed] [Google Scholar]
- Frank H., Nicholson G. J., Bayer E. Rapid gas chromatographic separation of amino acid enantiomers with a novel chiral stationary phase. J Chromatogr Sci. 1977 May 10;15(5):174–176. doi: 10.1093/chromsci/15.5.174. [DOI] [PubMed] [Google Scholar]
- Godal H. C., Abildgaard U. Gelation of soluble fibrin in plasma by ethanol. Scand J Haematol. 1966;3(5):342–350. doi: 10.1111/j.1600-0609.1966.tb02378.x. [DOI] [PubMed] [Google Scholar]
- Goding J. W. Antibody production by hybridomas. J Immunol Methods. 1980;39(4):285–308. doi: 10.1016/0022-1759(80)90230-6. [DOI] [PubMed] [Google Scholar]
- Hantgan R. R., Hermans J. Assembly of fibrin. A light scattering study. J Biol Chem. 1979 Nov 25;254(22):11272–11281. [PubMed] [Google Scholar]
- Hui K. Y., Haber E., Matsueda G. R. Monoclonal antibodies to a synthetic fibrin-like peptide bind to human fibrin but not fibrinogen. Science. 1983 Dec 9;222(4628):1129–1132. doi: 10.1126/science.6648524. [DOI] [PubMed] [Google Scholar]
- Kierulf P., Godal H. C. Fibrinaemia and multiple thrombi in pancreatic carcinoma. A case studied with quantitative N-terminal analysis. Scand J Haematol. 1972;9(4):370–376. doi: 10.1111/j.1600-0609.1972.tb00954.x. [DOI] [PubMed] [Google Scholar]
- Kierulf P., Godal H. C. Fibrinaemia and multiple thrombi in pancreatic carcinoma. A case studied with quantitative N-terminal analysis. Scand J Haematol. 1972;9(4):370–376. doi: 10.1111/j.1600-0609.1972.tb00954.x. [DOI] [PubMed] [Google Scholar]
- Kudryk B. J., Collen D., Woods K. R., Blombäck B. Evidence for localization of polymerization sites in fibrinogen. J Biol Chem. 1974 May 25;249(10):3322–3325. [PubMed] [Google Scholar]
- Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
- König W., Geiger R. Eine neue Methode zur Synthese von Peptiden: Aktivierung der Carboxylgruppe mit Dicyclohexycarbodiimid unter Zusatz von 1-Hydroxy-benzotriazolen. Chem Ber. 1970;103(3):788–798. doi: 10.1002/cber.19701030319. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Largo R., Heller V., Straub P. W. Detection of soluble intermediates of the fibrinogen-fibrin conversion using erythrocytes coated with fibrin monomers. Blood. 1976 Jun;47(6):991–1002. [PubMed] [Google Scholar]
- Lipinski B., Gurewich V., Nowak A., Wetmore R. The effect of heparin and dipyridamole on the deposition of fibrin-like material in rabbits infused with soluble fibrin monomer or fibrinogen. Thromb Res. 1974 Sep;5(3):343–358. doi: 10.1016/0049-3848(74)90172-8. [DOI] [PubMed] [Google Scholar]
- Lipinski B., Wegrzynowicz Z., Budzynski A. Z., Kopeć M., Latallo Z. S., Kowalski E. Soluble unclottable complexes formed in the presence of fibrinogen degradation products (FDP) during the fibrinogen-fibrin conversion and their potential significance in pathology. Thromb Diath Haemorrh. 1967 Feb 28;17(1-2):65–77. [PubMed] [Google Scholar]
- Mahn I., Müller-Berghaus G. Studies on catabolism of 125I-labelled fibrinogen in normal rabbits and in rabbits with indwelling intravenous catheters: methodologic aspects. Haemostasis. 1975;4(1):41–50. [PubMed] [Google Scholar]
- Marder V. J., Shulman N. R., Carroll W. R. High molecular weight derivatives of human fibrinogen produced by plasmin. I. Physicochemical and immunological characterization. J Biol Chem. 1969 Apr 25;244(8):2111–2119. [PubMed] [Google Scholar]
- Matthias F. R., Reinicke R., Heene D. L. Affinity chromatography and quantitation of soluble fibrin from plasma. Thromb Res. 1977 Mar;10(3):365–384. doi: 10.1016/0049-3848(77)90148-7. [DOI] [PubMed] [Google Scholar]
- Müller-Berghaus G., Róka L., Lasch H. G. Induction of glomerular microclot formation by fibrin monomer infusion. Thromb Diath Haemorrh. 1973 May 10;29(2):375–383. [PubMed] [Google Scholar]
- Niewiarowski S., Gurewich V. Laboratory identification of intravascular coagulation. The serial dilution protamine sulfate test for the detection of fibrin monomer and fibrin degradation products. J Lab Clin Med. 1971 Apr;77(4):665–676. [PubMed] [Google Scholar]
- Nossel H. L. Relative proteolysis of the fibrinogen B beta chain by thrombin and plasmin as a determinant of thrombosis. Nature. 1981 May 14;291(5811):165–167. doi: 10.1038/291165a0. [DOI] [PubMed] [Google Scholar]
- Nossel H. L., Yudelman I., Canfield R. E., Butler V. P., Jr, Spanondis K., Wilner G. D., Qureshi G. D. Measurement of fibrinopeptide A in human blood. J Clin Invest. 1974 Jul;54(1):43–53. doi: 10.1172/JCI107749. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Olexa S. A., Budzynski A. Z. Evidence for four different polymerization sites involved in human fibrin formation. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1374–1378. doi: 10.1073/pnas.77.3.1374. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SHAINOFF J. R., PAGE I. H. Significance of cryoprofibrin in fibrinogen-fibrin conversion. J Exp Med. 1962 Nov 1;116:687–707. doi: 10.1084/jem.116.5.687. [DOI] [PMC free article] [PubMed] [Google Scholar]
- THOMAS L., SMITH R. T., VON KORFF R. Cold-precipitation by heparin of a protein in rabbit and human plasma. Proc Soc Exp Biol Med. 1954 Aug-Sep;86(4):813–818. doi: 10.3181/00379727-86-21241. [DOI] [PubMed] [Google Scholar]