Characterization of Monoclonal Antibodies to Human Tissue-Type Plasminogen Activator: Catalytic Inhibition and One-Two Chain Discriminatory Reactivities

Summary

A new set of monoclonal antibodies was generated against the tissue plasminogen activator (t-PA). One of the antibodies, 1:3 C5, was found to be able to distinguish between the one- and two-chain form of t-PA and also exerted significant amidolytic inhibitory activity. Several of the antibodies, as judged from their binding properties in immunosorbent tests, were found to be suitable for immunoaffinity purification purposes, i.e. 1:3C5, 1:3 G5, and 1:2 B9. Three of the mabs, 1:2 B9, 1:3 G5 and 2:2 BIO, were selectively reactive with the A-chain of t-PA, whereas indirect evidences indicated 1:3 C5 to be reactive with a conformational epitope on the B-chain. Three of the antibodies were reactive with porcine t-PA. This new set of antibodies should prove useful for structure-function investigations of t-PA.

Keywords

Tissue plasminogen activator - Fibrinolysis - Monoclonal antibodies

• References

• 1 Collen D. On the regulation and control of fibrinolysis. Thromb Haemostas 1980; 43: 77-89
• 2 Rijken DC, Wijngaards G, Welbergen J. Relationship between tissue plasminogen activator and the activators in blood and vascular wall. Thromb Res 1980; 18: 815-830
• 3 Thorsen S, Glas-Greenwalt P, Astrup T. Differences in the binding to fibrin of urokinase and tissue plasminogen activator. Thromb Diath Haemorrh 1972; 28: 65-74
• 4 Wallén P. Structure and function of tissue plasminogen activator and urokinase. In: Fundamental and Clinical Fibrinolysis. Castellino et al. (eds) Academic Press; New York: 1987: 1-18
• 5 Hoylaerts M, Rijken DC, Collen D. Kinetics of the activation of plasminogen by human tissue plasminogen activator. Role of fibrin. J Biol Chem 1982; 257: 2912-2919
• 6 Rånby M. Studies on kinetics of plasminogen activation by tissue plasminogen activator. Biochim Biophys Acta 1982; 704: 461-469
• 7 Weimar W, Stibbe J, Vanseyen AJ, Billian A, Desomer P, Collen D. Specific lysis of an iliofemoral thrombus by administration of extrinsic (tissue-type) plasminogen activator. Lancet 1981; 2: 1018-1020
• 8 Korninger C, Matsuo O, Suy R, Stassen JM, Collen D. Thrombolysis with human extrinsic (tissue-type) plasminogen activator in dogs with femoral vein thrombosis. J Clin Invest 1982; 69: 573-580
• 9 Bergmann SR, Fox KA A, Terpogossian MM, Sobel BE, Collen D. Clot selective coronary thrombolysis with tissue type plasminogen activator. Science 1983; 220: 1181-1183
• 10 Van de Werf F, Bergmann SR, Fox KA, de Geest H, Hoyng CF, Sobel BE, Collen D. Coronary thrombolysis with intravenously administered human tissue-type plasminogen activator produced by recombinant DNA technology. Circulation 1984; 69: 605-610
• 11 Van de Werf F, Ludbrook PA, Bergmann SR, Tiefenbrunn AJ, Fox KA, de Geest H, Verstraete M, Collen D, Sobel B. Coronary thrombolysis with tissue-type plasminogen activator in patients with evolving myocardial infarction. N Eng J Med 1984; 310: 609-613
• 12 Wallen P, Rånby M, Bergsdorf N, Kok P. Purification and characterization of tissue plasminogen activator; on the occurrence of two different forms and their enzymatic properties. In: Progress in Fibrinolysis. Davidson JF, Nilsson IM, Asted B. (eds) (; 5). Churchill Livingstone; Edinburgh: 1981: 229-232
• 13 Pennica D, Holmes WE, Kohr WJ, Harkins RN, Vehar GA, Ward CA, Bennett WF, Yelverton E, Seeburg PH, Heyneker HL, Goeddel DV, Collen D. Cloning and expression of human tissue-type plasminogen activator cDNA in E.coli. . Nature 1983; 301: 214-221
• 14 Banyai L, Varadi A, Patthy L. Common evolutionary origin of the fibrin-binding structures of fibronectin and tissue-type plasminogen activator. FEBS Lett 1983; 163: 37-41
• 15 MacGregor IR, Micklem LR, James K, Pepper DS. Characterization of epitopes on human tissue plasminogen activator recognised by a group of monoclonal antibodies. Thromb Haemostas 1985; 53: 45-50
• 16 Holvoet P, Lijnen HR, Collen D. Characterization of functional domains in human tissue-type plasminogen activation with the use of monoclonal antibodies. Eur J Biochem 1986; 158: 173-177
• 17 Holvoet P, Boes J, Collen D. Measurement of free, one-chain, tissue- type plasminogen activator in human plasma with an enzymelinked immunosorbent assay based on an active site-specific murine monoclonal antibody. Blood 1987; 69: 284-289
• 18 Matsuo O, Tanhara Y, Okada K, Bundo H, Nagoya T. Monoclonal antibody to human tissue-plasminogen activator. Thromb Res 1984; 36: 517-526
• 19 Tahada A, Shizume K, Ozawa T, Takahashi S, Takada Y. Characterization of various antibodies against tissue plasminogen activator using highly sensitive enzyme immunoassay. Thromb Res 1986; 42: 63-72
• 20 Wallén P, Pohl G, Bergsdorf N, Rånby M, Ny T, Jornvall H. Purification and characterization of a melanoma cell plasminogen activator. Eur J Biochem 1983; 132: 681-686
• 21 Wallén P, Bergsdorf N, Rånby M. Purification and identification of two structural variants of porcine tissue plasminogen activator by affinity adsorption on fibrin. Biochim Biophys Acta 1982; 719: 318-328
• 22 Ichinose A, Takio K, Fujikawa K. Localization of the binding site of tissue-type plasminogen activator to fibrin. J Clin Invest 1986; 78: 163-169
• 23 Rijken DC, Groenevelt E. Isolation and functional characterization of the heavy and light chains of human tissue-type plasminogen activator. J Biol Chem 1986; 261: 3096-3102
• 24 Kearny J, Radbruch F, Liesegang B, Rajewsky K. A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. J Immunol 1979; 123: 1548-1550
• 25 Uotila M, Engvall E, Rouslahti E. Monoclonal antibodies to human alpha-feto protein. Mol Immunol 1980; 17: 791-794
• 26 Chalon MP, Milne RW, Vaerman IP. Interactions between mouse immunoglobulins and staphylococcal protein A. Scand J Immunol 1979; 9: 359-364
• 27 Carlsson L, Folkersen J, Stigbrand T. Antigenic determinants of pregnancy-associated α₂-glycoprotein and α₂-macroglobulin defined by poly- and monoclonal antibodies. Mol Immunol 1985; 22: 1073-1080
• 28 Lau HKF, Lui AY Y, Wong NS. Identification of plasmin as the major contaminant in immunoglobulin preparations. Comp Biochem Physiol 1986; 84B: 507-511
• 29 Norrman B, Pohl G, Jomvall H, Wallen P. Proteolytically induced variations in the enzymatic properties of tissue plasminogen activator. Eur J Biochem 1986; 159: 7-13
• 30 Rånby M, Suensen E, Bergsdorf N, Norrman B, Wallen P. Tissue plasminogen activator kinetics. Prog Fibrinolys 1983; 6: 182-184
• 31 Rånby M, Norrman B, Wallen P. A sensitive assay for tissue plasminogen activator. Thromb Res 1982; 77: 743-749
• 32 Norrman B, Wallén P, Rånby M. Fibrinolysis mediated by tissue plasminogen activator: Disclosure of a kinetic transition. Eur J Biochem 1985; 149: 193-200
• 33 Deutsch DG, Mertz ET. Plasminogen: Purification from human plasma by affinity chromatography. Science 1970; 170: 1095-1096
• 34 Wallén P, Wiman B. Characterization of human plasminogen. Biochim Biophys Acta 1972; 257: 122-134
• 35 Norrman BA, Frangsmyr LC, Stigbrand T, Wallen PB. A monoclonal antibody to tissue plasminogen activator with affinity for the Kringle regions: Implications for fibrin affinity and enzymatic mechanism. Fibrinolysis 1987; 1: 135-142
• 36 Liu CY, Handley DA, Wallen P, Smith J. Fibrin potentiation of plasminogen activation by tissue plasminogen activator on bovine endothelial cell surface. Abstract No. 98 The 8th International Congress on Fibrinolysis; Vienna: 1986
• 37 Wallén P, Pohl G, Bergsdorf N, Rånby M, Jörnvall H. Purification and characterization of a melanoma cell plasminogen activator. Eur J Biochem 1983; 132: 681-686
• 38 van Zonneveld AJ, Veerman H, Brakenhoff JP J, Aarden LA, Cajot F, Pannekoek H. Mapping of epitopes on human tissue-type plasminogen activator with recombinant deletion mutant proteins. Thromb Haemostas 1987; 57: 82-86
• 39 Wallén P, Bergsdorf N, Pohl G, Stigbrand T. Differentiation between one-chain and two-chain tissue type plasminogen activator by monoclonal antibodies. Haemostasis 1984; 14: 42