Thromb Haemost 1992; 68(03): 273-277
DOI: 10.1055/s-0038-1656364
Original Article
Schattauer GmbH Stuttgart

A Rapid Monoclonal Antibody-Based Enzyme Immunoassay (EIA) for the Quantitative Determination of Soluble Fibrin in Plasma

W Nieuwenhuizen
The IVVO-TNO, Gaubius Laboratory, Leiden, The Netherlands
,
E Hoegee-De Nobel
The IVVO-TNO, Gaubius Laboratory, Leiden, The Netherlands
,
R Laterveer
The IVVO-TNO, Gaubius Laboratory, Leiden, The Netherlands
› Institutsangaben
Weitere Informationen

Publikationsverlauf

Received 31. Januar 1992

Accepted after revision 30. März 1992

Publikationsdatum:
04. Juli 2018 (online)

Summary

Soluble fibrin is considered as a molecular marker for intravascular fibrin formation, and impending thrombotic events. Most of the existing assays are less suitable for routine clinical applications and their specificity may be limited.

We have developed a sandwich-type EIA with a fibrin-specific MoAb described by us before (Proc Natl Acad Sci 1989; 86: 8951) as the capture antibody. An other MoAb (G8; Thromb Haemostas 1988; 60: 415) with an epitope in the carboxyl-terminal sections of the fibrin α-chains, was labeled with peroxidase and used as the tagging antibody. The EIA is calibrated against plasma spiked with known concentrations of soluble fibrin. The time-to-result of the EIA is only 1.5 h. Concentrations as low as 0.5 µg soluble fibrin/ml plasma are readily measurable. Heparin has no effect on the results. Fibrinogen and fibrin(ogen) degradation products are not detected. The values of fibrinopeptide A and soluble fibrin values found with the “COA-SET soluble fibrin” assay correlated well with the soluble fibrin values found with our EIA i.e. r = 0.998 and 0.984, respectively. The run-to-run variabilities were 7.9% and 6.6% for samples with low and high soluble fibrin concentrations, respectively. The within-run variabilities were 2.5, 1.8, 4.0 and 4.6% for samples with 1, 0.5, 0.25 and 0.125 µg soluble fibrin/ml, respectively.

The sensitivity, specificity, accuracy and short time-to-result make our EIA suitable for routine clinical applications and the monitoring of the effectivity of heparinization.

 
  • References

  • 1 Nieuwenhuizen W. The use of monoclonal antibodies in demonstrating different fibrinogen derivatives. In: Müller-Berghaus G, Scheefers-Borchel U, Selmayer E, Henschen A. (eds) Excerpta Medica: Amsterdam: 1986. pp 245-246
  • 2 Nieuwenhuizen W. Plasma assays of fibrinogen/fibrin degradation products and their clinical relevance. Fibrinogen 2. Biochemistry, Physiology and Clinical Relevance Lowe GDO, Douglas JT, Forbes CD, Henschen A. Excerpta Medica, Amsterdam: 1987: 173-80
  • 3 Nieuwenhuizen W. New strategies in the determination of fibrin and fibrin(ogen) derivatives by monoclonal antibodies. Blut 1988; 57: 285-291
  • 4 Doolittle RF. Structural aspects of the fibrinogen to fibrin conversion. Adv Protein Chem 1973; 27: 1-109
  • 5 Doolittle RF. Fibrinogen and fibrin. Sci Am 1981; 245: 092-101
  • 6 Shainoff JR, Page JH. Significance of cryoprofibrin in fibrinogen-fibrin conversions. J Exp Med 1962; 116: 687-707
  • 7 Brass EP, Forman WB, Edwards RV, Lindan O. Fibrin formation: The role of the fibrinogen-fibrin complexes. Thromb Haemostas 1976; 36: 36-48
  • 8 Graeff H, Hafter R, Von Hugo R. On soluble fibrinogen-fibrin complexes. Thromb Res 1979; 16: 575-576
  • 9 Hermans J, McDonagh J. Fibrin: Structure and interactions. Semin Thromb Hemostas 1982; 8: 11-24
  • 10 Bick RL. Disseminated intravascular coagulation: A clinical/laboratory study of 48 patients. Ann N Y Acad Sci 1981; 370: 843-855
  • 11 Siegal T, Seligsohn U, Aghai E, Modan M. Clinical and laboratory aspects of disseminated intravascular coagulation (DIC): A study of 118 cases. Thromb Haemostas 1978; 39: 122-140
  • 12 Colman RW, Robboy SJ, Minna JD. Disseminated intravascular coagulation (DIC): An approach. Am Rev Med 1972; 52: 679-689
  • 13 Thomas DP, Niewiarowski S, Myers AR, Block KJ, Colman RW. A comparative study of four methods for detecting fibrinogen degradation products in patients with various diseases. N Engl J Med 1970; 238: 663-668
  • 14 Spero JA, Lewis JH, Hasiba U. Disseminated intravascular coagulation: Findings in 346 patients. Thromb Haemostas 1980; 38: 28-33
  • 15 Okajima K, Koga S, Okabe H, Inoue M, Takatsuki K. Characterization of the fibrinolytic state by measuring stable cross-linked fibrin degradation products in disseminated intravascular coagulation associated with acute promyelocytic leukemia. Acta Haematol 1989; 81: 15-18
  • 16 Fruchtman S, Aledort LM. Disseminated intravascular coagulation. J Am Coll Cardiol 1986; 8: 159-167
  • 17 Breen FA, Tullis JL. Ethanol gelation: A rapid screening test for intravascular coagulation. Ann Intern Med 1968; 69: 1197-1206
  • 18 Godal HC, Abildgaard U. Gelation of soluble fibrin in plasma by ethanol. Scand J Haematol 1966; 3: 342-350
  • 19 Gurewich V, Hutchinson E. Detection of intravascular coagulation by a serial dilution protamine sulfate test. Ann Intern Med 1971; 75: 895-905
  • 20 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; 77: 665-676
  • 21 Watanabe K, Tullis JL. Precipitation of fibrin monomers and fibrin degradation products by ristocetin. Am J Med Sci 1978; 275: 337-344
  • 22 Watanabe K, Tullis JL. Ristocetin precipitation test: A new simple test for the detection of fibrin monomer and fibrin degradation products. Am J Clin Pathol 1978; 70: 691-696
  • 23 Largo R, Heller V, Straub PW. Detection of soluble intermediates of the fibrinogen-fibrin conversion using erythrocytes coated with fibrin monomers. Blood 1976; 47: 0991-1002
  • 24 Fletcher AP, Alkjaersig N, O’Brien J, Tulevski VG. Blood hypercoagulability and thrombosis. Transact Assoc Am Physicians 1970; 83: 159-167
  • 25 Graeff H, von Hugo R. Identification of fibrinogen derivatives in plasma samples. Thromb Diath Haemorrh 1972; 27: 610-618
  • 26 Heene DL, Matthias FR. Adsorption of fibrinogen derivatives on insolubilized fibrinogen and fibrin monomer. Thromb Res 1973; 2: 137-154
  • 27 Matthias FR, Reinicke R, Heene DL. Affinity chromatography and quantification of soluble fibrin from plasma. Thromb Res 1977; 10: 365-384
  • 28 Wiman B, Rånby M. Determination of soluble fibrin in plasma by a rapid and quantitative spectrophotometric assay. Thromb Haemostas 1986; 55: 189-193
  • 29 Scheefers-Borchel U, Müller-Berghaus G, Fuhge P, Eberle R, Heimburger N. Discrimination between fibrin and fibrinogen by a monoclonal antibody against a synthetic peptide. Proc Natl Acad Sci USA 1985; 82: 7091-7095
  • 30 Schielen WJG, Voskuilen M, Tesser GI, Nieuwenhuizen W. The sequence Aα-[148–160] in fibrin, but not in fibrinogen, is accessible to monoclonal antibodies. Proc Natl Acad Sci USA 1989; 86: 8951-8954
  • 31 Hoegee-De Nobel E, Voskuilen M, Briët E, Brommer EJP, Nieuwenhuizen W. A monoclonal antibody-based quantitative enzyme immunoassay for the determination of plasma fibrinogen concentrations. Thromb Haemostas 1988; 60: 415-418
  • 32 Bouvet JP, Pires R, Pillot J. A modified gel filtration technique producing an unusual exclusion volume of IgM: a simple way of preparing monoclonal IgM. J Immunol Meth 1984; 66: 299-305
  • 33 Ey PL, Prowse SJ, Jenkin CR. Isolation of pure IgG1, IgG2a and IgG2b immunoglobulins from mouse serum using protein A Sepharose. Immunochemistry 1978; 15: 429-436
  • 34 Carlsson J, Orevin H, Axén R. Protein thiolation and reversible protein-protein conjugation. N-Succinimidyl 3-(2-pyridyldithio) propionate: a new heterobifunctional reagent. Biochem J 1978; 173: 723-737
  • 35 Nieuwenhuizen W, Creighton LC, Gaffney PJ, Graeff H, Hafter R, Hoegee-De Nobel E, Müller-Berghaus G, Scheefers-Borchel U, Thurmayer R, Davies JA, Duckert F, Jespersen J, Matthias FR, Prentice CRM, Preston FE, Samama MM. A double blind comparative study of six monoclonal antibody-based plasma assay for fibrinogen derivatives. Fibrinogen 2. Biochemistry Physiology and Clinical Relevance Lowe GDO, Douglas JT, Forbes CD, Henschen A. Excerpta Medica, Amsterdam: 1987: 181-188
  • 36 Koopman J, Haverkate F, Koppert P, Nieuwenhuizen W, Brommer EJP, Van der Werf WGC. New enzyme immunoassay of fibrinfibrinogen degradation products in plasma using a monoclonal antibody. J Lab Clin Med 1987; 109: 75-84