Immunodetection of Human Fibrin Using Monoclonal Antibody-64C5 in an Extracorporeal Chicken Model

Summary

The potential use of a fibrin-specific monoclonal antibody-64C5 as a thrombus detecting agent was examined in a quantifiable extracorporeal circulatory model using the chicken. This species was selected because antibody-64C5 did not cross-react with chicken fibrin. Human fibrinogen was clotted with human thrombin and factor XIII on filter paper disks, which were then inserted into an ex-vivo chamber that interrupted an extracorporeal loop between a carotid artery and a jugular vein. Antibody-64C5 was injected intravenously and after an equilibration period, disks bearing human fibrin were placed in the extracorporeal chamber and exposed to circulating antibody. Uptake which was quantified by treating the disks with ¹²⁵I-goat antimouse-(Fab’)₂ was rapid, with 70 percent of maximal uptake occurring within 10 min. Fibrinolysis, although measurable, had no adverse effect upon uptake of the monoclonal antifibrin-64C5.

Keywords

Fibrin-specific monoclonal antibody - Chicken model - Extracorporeal circulation - Thrombus detection

• References

• 1 Laffel GL, Braunwald E. Thrombolytic therapy: a new strategy for the treatment of acute myocardial infarction. N Eng J Med 1984; 311: 710-717 ibid. 770-6.
• 2 Gurewich V, Pannell R, Louie S, Kelley P, Suddith RL, Greenlee R. Effective and fibrin-specific clot lysis by a zymogen precursor form of urokinase (pro-urokinase). J Clin Invest 1984; 73: 1731-1739
• 3 Bergmann SR, Fox KA, Ter-Pogossian MM, Sobel BE, Collen D. Clot-selective coronary thrombolysis with tissue-type activator. Science 1983; 220: 1181-1183
• 4 Kakkar VV, Nicolaides AN, Renney JT, Friend JR, Clarke MB. ¹²⁵I-labelled fibrinogen test adapted for routine screening for deep-vein thrombosis. Lancet 1970; 01: 540-542
• 5 Thakur ML, Welch MJ, Joist JH, Coleman RE. Indium-111 labeled platelets: Studies on preparation and evaluation of in-vitro and in-vivo functions. Thromb Res 1976; 09: 345-357
• 6 Kwaan HC, Grumet G. Clinical use of ⁵¹Cr-leukocytes in detection of deep vein thrombosis. Circulation 1972; 46 (Suppl. 02) II-52
• 7 Knight LC, Olexa SA, Malmud LS, Budzynski AZ. Specific uptake radioiodinated fragment E1 by venous thrombi in pigs. J Clin Invest 1983; 72: 2007-2013
• 8 Gomez RL, Wheeler HB, Belko JS. et al. Observations of the uptake of a radioactive fibrinolytic enzyme by intravascular clots. Ann Surg 1963; 158: 905-911
• 9 Harwig SL, Harwig JF, Sherman LA, Coleman RE, Welch MJ. Radioiodinated plasminogen: An imaging agent for pre-existing thrombi. J Nucl Med 1977; 18: 42-45
• 10 Deacon JM, Ell PJ, Anderson P, Khan O. Technetium (99m)-plasmin: a new test for the detection of deep vein thrombosis. Br J Radiol 1980; 53: 673-677
• 11 Gross R. Findings with labeled streptokinase in-vitro and in-vivo. Proceedings of the Ninth Congress of the European Society of Haematology, Lisbon, 1963, Basel, Karger S. 1963: 1342-1435
• 12 Rhodes BA, Bell WR, Malmud LS. et al. Labelling and testing of urokinase and streptokinase – New tracers for the detection of thromboemboli, in Radiopharmaceuticals and Labelled Compounds, Vol 2, Vienna, IAEA. 1973: 163-169
• 13 Spar IL, Perry JM, Benz LL, DeWeese JA, Mahoney EB. Detection of left atrial thrombi: Scintillation scanning after administration of ¹³¹I-rabbit antibodies to human fibrinogen. Am Heart J 1969; 78: 731-739
• 14 Hui KY, Haber E, Matsueda GR. Monoclonal antibodies to a synthetic fibrin-like peptide bind to human fibrin but not fibrinogen. Science 1983; 222: 1129-1132
• 15 Walker L, Catlin A. A simplified method for preparation of fibrinogen. Thrombos Diathes Haemorrh 1971; 26: 99-102
• 16 Chen R, Doolittle RF. Isolation, characterisation and location of a donor-receptor unit from cross-linked fibrin. Proc Natl Acad Sci USA 1970; 66: 472-479
• 17 McKee PA, Mattock P, Hill RL. Subunit structure of human fibrinogen, soluble fibrin and cross-linked insoluble fibrin. Proc Natl Acad Sci USA 1970; 66: 738-744
• 18 Greenwood FC, Hunter WM, Glover JS. The preparation of ¹³¹I-labelled human growth hormone of high specific radioactivity. Biochem J 1963; 89: 114-123
• 19 Collen D. On the regulation and control of fibrinolysis. Thromb Haemostas 1980; 43: 77-89
• 20 Robbins KC, Summaria L. In: Methods in Enzymology. Perlmann GE, Lorand L. (Eds). Academic Press; New York: 1970. Vol XIX: 184-199
• 21 Hawkey CM. Fibrinolysis in Animals. Symp Zool Soc London 1970; 27: 133-150
• 22 Niewiarowski S, Latallo Z. Comparative studies of the fibrinolytic system of sera of various vertebrates. Thrombos Diathes Haemorrh 1959; 3: 404-417
• 23 Kaminski M, MacDonagh J. Studies on the mechanism of thrombin interaction with fibrin. J Biol Chem 1983; 258: 10530-10535