Abstract
The detection of thrombi in rabbits has been investigated with131I-labelled DD-3136/22, a monoclonal antibody (Mab) reactive at high affinity (Kd=2.68 x 10−10 M) with human D Dimer (DD). DD-3136/22 bound well to both “fresh” and “aged” human clots in an in vitro assay but showed poor binding to rabbit clots. However, reactivity was restored to rabbit blood if it was seeded, before clotting, with human DD covalently coupled to Sepharose beads. Thus, a rabbit model was developed in which blood was allowed to clot around DD-Sepharose beads introduced into the jugular vein. Gamma camera imaging showed that intact131I-labelled DD-3136/22 localised to these clots within 24 h. Uptake at this time was 0.202±0.012% injected dose per gram (%ID/g) compared with 0.086±0.018%ID/g after injection of control antibody.131I-labelled F(ab′)2, fragments of DD-3136/22 allowed earlier scintigraphic detection of the clot which was evident 4 h after injection. Uptake in the clot at 24 h was 0.154±0.038 %ID/g compared with 0.109±0.027 %ID/g for a control F(ab')2. As antigen levels in the clot are estimated to be less than 300 μg DD, thus representing a very small human clot, the DD-3136/22 Mab would appear to have a good potential for the sensitive detection of thrombi in a clinical setting.
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References
Bini A, Fenoglio J Jr, Sobel J, Owen J, Fejgl M, Kaplan KI (1987) Immunochemical characterization of fibrinogen, fibrin I and Fibrin II in human thrombi and atherosclerotic lesions. Blood 69:1038–1045
Bode C, Matsueda GR, Hui KY, Haber E (1985) Antibody-directed urokinase: a specific fibrinolytic agent. Science 229:765–767
Bosnjakovic VB, Jankovic BD, Horvat J, Cvoric J (1977) Radiolabelled anti-human fibrin antibody: a new thrombus-detecting agent. Lancet I:452–454
Bosnjakovic V, Jankovic BD, Horvat J, Nastic-Miric D, Djukic V, Pavlovic S (1988) The validity of radiolabelled anti fibrin antibody for deep vein thrombosis imaging. Eur J Nucl Med 14:489–494
Brenner B, Marder VJ, Francis CW (1988) D-dimer immunoreactivity of digests of soluble fibrin polymers and of fibrin clots depends on the extent of degradation. Blood [Suppl 1] 72:363a
Buraggi GL (1986) Radioimmunodetection of malignant melanoma with the 225.285 monoclonal antibody to HMW-MAA. Nucl Med 25:220–224
Feitsma RIJ, Blok D, Wasser MNJM, Nieuwenhuizen W, Pauwels EKJ (1987) A new method for99Tcm-labelling of proteins with an application to clot detection with an antifibrin monoclonal antibody. Nucl Med Comm 8:771–777
Francis CW, Marder VT (1986) Concepts of clot lysis. Ann Rev Med 37:187–204
Griswold WR, Nelson DP (1985) Calculation of monoclonal antibody affinity constants directly from antibody dilution curves. Immunol Lett 9:15–18
Houranieh A, Statland BE, Davis GL, Ito RK, Relias V, Kudryk B, Yunis EJ (1988) Monoclonal antibody to human cross-linked fibrin. Hybridoma 7:55–68
Khaw BA, Strauss HW, Yasuda T, Hui K, Fallon JT, Haber E, Matsueda G (1985) In-111 labeled monoclonal anti-(fibrin specific) antibody: detection of pulmonary emboli. J Nucl Med 26:P21
Knight LC, Maurer AH, Ammar IA, Shealy DJ, Mattis JA (1988) Evaluation of indium-111-labeled anti-fibrin antibody for imaging vascular thrombi. J Nucl Med 29:494–502
Lamoyi E, Nisonoff A (1983) Preparation of F(ab')2 fragments from mouse IgG of various subclasses. J Immunol Methods 56:235–243
Liau CS, Haber E, Matsueda GR (1987) Evaluation of monoclonal antifibrin antibodies by their binding to human blood clots. Thromb Haemost 57:49–54
Nieuwenhuizen W (1986) The use of monoclonal antibodies in demonstrating different fibrinogen derivatives. In: Muller Berghaus G et al. (eds) Fibrinogen and its derivatives. Elsevier, Amsterdam, p 245
Oster ZH, Srivastava SC, Som P, Meinken GE, Scudder LE, Yamamoto K, Atkins HL, Brill AB, Coller BS (1985) Thrombus radioimmunoscintigraphy: an approach using monoclonal antiplatelet antibody. Proc Natl Acad Sci USA 82:3465–3468
Peters AM, Lavender JP, Needham SG, Loutfi I, Snook D, Epenetos AA, Lumley P, Keery RJ, Hogg N (1986) Imaging thrombus with radiolabelled monoclonal antibody to platelets. Br Med J (Clin Res) 293:1525–1527
Rosebrough SF, Kudryk B, Grossman ZD, McAfee JG, Subramanian G, Ritter-Hrncirik CA, Witanowski LS, Tillapaugh-Fay G (1985) Radioimmunoimaging of venous thrombi using iodine-131 monoclonal antibody. Radiology 156:515–517
Rosebrough SF, Grossman ZD, McAfee JG, Kudryk B, Subramanian G, Ritter-Hrncirik CA, Witanowski LS, Tillapaugh-Fay G, Urrutia E (1987) Aged venous thrombi: radioimmunoimaging with fibrin-specific monoclonal antibody. Radiology 162:575–577
Rosebrough SF, Grossman ZD, McAfee JG, Kudryk BJ, Subramanian G, Ritter-Hrncirik CA, Witanowski LS, Tillapaugh-Fay G, Urrutia E, Zapf-Longo C (1988) Thrombus imaging with indium-111 and iodine-131-labeled fibrin-specific monoclonal antibody and its F(ab')2 and Fab fragments. J Nucl Med 29:1212–1222
Rowbotham BJ, Carroll P, Whitaker AN, Bunce IH, Cobcroft RG, Elms MJ, Masci PP, Bundesen PG, Rylatt DB, Webber AJ (1987) Measurement of crosslinked fibrin derivatives — use in the diagnosis of venous thrombosis. Thromb Haemost 57:59–61
Rylatt DB, Blake AS, Cottis LE, Massingham DA, Fletcher WA, Masci PP, Whitaker AN, Elms M, Bunce I, Webber AJ, Wyatt D, Bundesen PG (1983) An immunoassay for human D dimer using monoclonal antibodies. Thromb Res 31:767–778
Schnee JM, Runge MS, Matsueda GR, Hudson NW, Seidman JG, Haber E, Quertermous T (1987) Construction and expression of a recombinant antibody-targeted plasminogen activator. Proc Natl Acad Sci USA 84:6904–6908
Som P, Oster ZH, Zamora PO, Yamamoto K, Sacker DF, Brill AB, Newell KD, Rhodes BA (1986) Radioimmunoimaging of experimental thrombi in dogs using technetium-99m-labeled monoclonal antibody fragments reactive with human platelets. J Nucl Med 27:1315–1320
Spar IL, Goodland RL, Schwartz SI (1965) Detection of preformed venous thrombi in dogs by means of I131-labeled antibodies to dog fibrinogen. Circ Res 17:322–329
Stuttle AWJ, Peters AM, Loutfi I, Lumley P, George P, Lavender JP (1988) Use of an anti-platelet monoclonal antibody F(ab')2 fragment for imaging thrombus. Nucl Med Comm 9:647–655
Walker KZ, Seymour-Munn K, Keech FK, Axiak SM, Bautovich GJ, Morris JG, Basten A (1986) A rat model system for radioimmunodetection of kappa myeloma antigen on malignant B cells. Eur J Nucl Med 12:461–467
Walker KZ, Khafagi F, Bautovich GJ, Boniface GR, Bundesen PG, Rylatt DB (1988a) Anti-fibrin monoclonal antibodies for radioimmunodetection: preliminary assessment in a rat model system. Thromb Res 52:269–278
Walker KZ, Seymour-Munn K, Towsin JE, Axiak SM, Raison RL, Bautovich GJ, Morris J, Basten A (1988b) Radioimmunolocalization and selective delivery of radiation in a rat model system: comparison of intact and fragmented antibody. Nucl Med Comm 9:517–526
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Walker, K.Z., Milner, L.J., Bautovich, G.J. et al. Detection of experimental thrombi in rabbits with an131I-labelled fibrin-specific monoclonal antibody. Eur J Nucl Med 16, 787–794 (1990). https://doi.org/10.1007/BF00833012
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DOI: https://doi.org/10.1007/BF00833012