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Radiolabelled monoclonal antibodies (McAb): An alternate approach to the conventional methods for the assessment of cardiomyocyte damage in an experimental brain-death pig model

Abstract

The present study was carried out to determine the possible use of cTn-I in the cardiac myofibrillar architecture, as a potential target forin vivo radioimmunodetection of cardiac damage in a brain death pig model. Radioiodiantion of the anti-cTn-I 5F4 McAb was carried out by lactoperoxidase method. The percentage iodine incorporation achieved was 70∼75%. The radioiodinated McAbs were purified on Sephadex G-25 column and characterised by Paper chromatography, Phast Gel electrophoresis and electroimmunoblotting. Radioiodinated anti-cTn-I 5F4 McAbs were employed alongside Pyrophosphate (Tc99m-PPi) and Thallium201 chloride (Tl201) in 24 landrace pigs (brain-dead=18 & sham-operated=6). The percentage cardiac uptake of the radiolabelled antibody injected dose was significantly higher in the brain dead animals (0.196%) as compared to that of sham-operated animals (0.11%). Specificin vivo localization of radiolabelled McAbs in the infarcted cardiac tissue was confirmed by computer-aided reconstruction of 3-D images of the isolated heart. The preliminary results of the study revealed preferential uptake of radiolabelled antibody at the site of myocyte damage resulting from artificially induced brain death.

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References Cited

  1. Beller, G. A., Khaw, B. A., Haber, E. and Smith, T. W. Localization of radiolabelled cardiac myosin specific antibody in myocardial infarct: comparison with Tc99m-Sn-PPi.Circulation, 55, 74–78 (1977).

  2. Bodor, G., Porter, S. and Ladenson J. H. Human cardiac troponin-I measurement in suspected MI with a double monoclonal antibody sandwich ELISA.Clin. Chem., 36, 1103 (1990).

  3. Bodor, G., Porter, S., Landt, Y. and Ladenson J. H. Development of monoclonal antibodies for an assay of cardiac troponin-I and preliminary results in the suspected cases of myocardial infarction.Clin. Chem., 38, 2203–14 (1992).

  4. Caputo, V., DeNardo, D., Antolini, M., Pittuco, G., Gallo, P., Scibilia, G., Macchiarelli, A. G., Cassisi, A., Caretta, Q., Bianco, G. and Marino, B. Myocardial necrosis imaging by In111-monoclonal antimyosin Fab.Nucl. Med. Biol. Int. J. Appl. Radioisot (Part-B), 16, 641–43 (1989).

  5. Corney, P. L., Rogers, P. L. and Johnson, D. K. Dual Isotope study of I125 and In111 labelled antibody in athymic mice.J. Nucl. Med., 30, 374–84 (1989).

  6. Cummins, B., Russell, G. J., Chandler, S. T., Pears, D. and Cummins, P.,In vivo imaging of the damaged cardiac muscle using radiolabelled cardiac troponin-I antibodies.J. Muscle Res. Cell Motility, 10, 162 (1989).

  7. Cummins, B., Russell, G. J., Chandler, S. T., Pears, D. and Cummins, P. Uptake of the radiolabelled cardiac specific Tn-I antibodies in myocardial infarction.Cardiovasc. Res., 24, 317–27 (1990).

  8. Cummins, P. and Perry, S. V. Troponin-I from human skeletal and cardiac muscles.Biochem. J., 171, 251–59 (1978).

  9. Cummins, P., McGurk, B. and Littler, W. A. Possible diagnostic use of cardiac specific contractile proteins in assessing cardiac damage.Clin. Sci., 56, 30 (1979)

  10. Dhoot, G. K., Gell, P. G. H. and Perry, S. V. The localization of the different forms of Tn-I in skeletal and cardiac muscle cells.Exp. Cell Res., 117, 357–70 (1978).

  11. Ebashi, S. Separation of troponin into three components.J. Biochem., 72, 787–90 (1972).

  12. Eckelman, W. C. and Paik, C. H. Labelling of antibodies with metals using bifunctional chelates. Zalutsky, M. R.. (Ed)Antibodies in Radiodiagnosis and Therapy. CRC Press, Florida, 103–128 (1989).

  13. Editorial. Hot spot imaging for acute myocardial imaging.J. Nucl. Med., 31, 143–45 (1990).

  14. Editorial: Troponin-T and myocardial damage.Lancet, 338, 23–24 (1991).

  15. Gibson, R. S. and Watson, D. D. Clinical applications of myocardial perfusion scintigraphy with Thallium201. Yu, P. N., & Goodwin, J. F. (Eds)Progress in Cardiology. Lea & Febiger, Philadelphia, 12, 67–107 (1983).

  16. Haider, Kh. H. and Stimson, W. H. Production and chacracterization of anti-cardiac troponin-I monoclonal antibodies.Disease Markers, 12, 187–97 (1994).

  17. Haider, Kh. H. and Stimson, W. H. Bovine cardiac troponin-I specific monoclonal antibodies which show species cross reactivity.Exp Mol. Med., 28, 71–76 (1996).

  18. Haider, Kh. H. and Stimson, W. H. Radioiodination and characterization of cTn-I specific monoclonal antibodies for cardiac imaging.Chinese J. Pharm. Sci., 48, 321–30 (1996).

  19. Hoberg, E., Eisenhut, M., Hoffman, M., Schuler, G., Brachman, J., Aidonidis, I., Natchkamp, J., Kubler, W. and Katus, H. A. Monoclonal antibodies specific for human cardiac myosin: selection, characterization and experimental infarction imaging.Eur. Heart J., 9, 328–36 (1988).

  20. Hunkler, N. M., Kullman, J. and Murphy, A. M. Troponin-I isoform expression in the human heart.Cir. Res., 69, 1409–1414 (1991).

  21. Katagiri, T., Koybayasji, Y., Sasai, Y., Toba, K. and Niitani, H. Alterations in the troponin subunits in myocardial infarction.Jap. Heart J., 22, 653–64 (1981).

  22. Ladenson, J. H., Porter, S., Landt, Y. and Bodor, G. Development and characterization of monoclonal antibody specific to troponin-I.Clin. Chem., 36, 1019 (1990).

  23. Lammeli, U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature, 227, 680–88 (1970).

  24. Larue, C., Defacque-Lacquement, H., Calzolari, C., Nguyen, D. L. and Pau, B. New monoclonal antibodies as probes for human cardiac troponin-I: Epitopic analysis with synthetic peptides.Mol. Immunol., 29, 271–78 (1992).

  25. Novitzky, D., Wicomb, W. N., Cooper, D. C. K., Rose, A. G. and Reichart, B. Prevention of myocardial injury during brain death by total cardiac sympathectomy in chacma baboon.Ann. Thorac. Surg., 41, 520–24 (1986).

  26. Novitzky, D., Wicomb, W. N., Cooper, D. C. K. and Tjaalgard, M. A. Improved cardiac function following hormonal therapy in brain dead pigs: relevance to organ donation.Cryobiology, 24, 1–10 (1987).

  27. Novitzky, D., Cooper, D. C. K. and Wicomb, W. N. Endocrine changes and metabolic response.Transplant. Proc., suppl. 7, 33–38 (1988).

  28. Russell, G., Cummins, B. and Cumins, P. Characterization of monoclonal antibodies to primate cardiac troponin-I.J. Muscle Res. Cell Motility, 10(2), 155 (1989).

  29. Saha, G. B., Go, R. T. and MacIntyre, W. J., Radio-pharmaceuticals for cardiovascular, imaging.Nucl. Med. Biol. Int. J. Radiat. Appl. Instrum. Part-B, 19, 1–20 (1992).

  30. Tamaki, N., Yamada T. and Matsumori, A. Indium-111antimyosin antibody imaging for detecting different stages of myocardial imaging: comparison with Tc99m-PPi imaging.J. Nucl. Med., 31, 136–42 (1990).

  31. Towbin, H., Staehelin, T. and Grdon, J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and application.Proc. Natl. Acad. Sci., 76, 435–54 (1979).

  32. Vallins, W. J., Brand, N. J., Dabhade, N., Butler-Brown, G., Yacoub, M. H. and Barton, P. J. R. Molecular cloning of human cTn-I using polymerase chain reaction.FEBS Letters, 17, 83–90 (1990).

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Correspondence to Kh. H. Haider.

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Haider, K.H., Stimson, W.H. Radiolabelled monoclonal antibodies (McAb): An alternate approach to the conventional methods for the assessment of cardiomyocyte damage in an experimental brain-death pig model. Arch. Pharm. Res. 21, 496–502 (1998). https://doi.org/10.1007/BF02975364

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Key words

  • Brain death
  • Cardiac Troponin-1
  • Monoclonal antibodies
  • Radioimmunoimaging
  • Radioiodination