Subscribe to RSS
DOI: 10.1055/s-0038-1656316
Screening for Fibrin Specific Monoclonal Antibodies: The Development of a New Procedure
Publication History
Received 01 November 1991
Accepted after revision 17 February 1992
Publication Date:
03 July 2018 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/199201/lookinside/thumbnails/10-1055-s-0038-1656316-1.jpg)
Summary
The acquisition of monoclonal antibodies specific for human fibrin has been impaired by the similarity in chemical composition between fibrinogen and fibrin and the conformational difference between immobilised and soluble fibrinogen. Five monoclonal antibodies (mabs) with a known affinity for fibrin have been subjected to screening procedures which involved the presentation of different forms of both fibrinogen and fibrin to the test mabs. It was observed by scanning electron microscopy that dried fibrin (denoted fibrin D), immobilised on the wells of PVC plates was morphologically similar to the fibrin found in human clots whereas PVC-immobilised fibrin monolayers (fibrin M) and a homogenised form of fibrin (fibrin FF) presented two very different morphological appearances. It was shown that lack of cross reactivity of a mab with an antigen (e.g. fibrinogen) was validly demonstrated only when both mab and antigen were present in the soluble state. These findings have allowed the generation of a screening procedure which involves the use of fibrin D on PVC plates in conjunction with whole human plasma incubated with the test antibody. This screening procedure has been validated using two mabs, one of which has an exclusive fibrin affinity while the other has a broad spectrum crossreactivity with both fibrinogen and fibrin. This procedure would ensure the acquisition of all the five fibrin-specific mabs used in this study while other less reliable screening procedures might not.
-
References
- 1 Kakkar VV, Nicolaides AN, Renney JTG, Friend JR, Clarke MB. I-labelled fibrinogen test adapted for routine screening for deep vein thrombosis. Lancet 1970; 1: 540-542
- 2 Deacon JM, Anderson P, Kahn O. Technesium99m-plasmin. A new test for the detection of deep vein thrombosis. Br J Radiol 1980; 53: 673
- 3 Knight LC, Primeau JL, Siegel BA, Welch MJ. Comparison of In-III labelled platelets and iodinated fibrinogen for the detection of deep vein thrombosis. J Nucl Med 1978; 19: 891-894
- 4 Kudryk B, Rohoza A, Ahadi M, Chin J, Wiebe ME. Specificity of a monoclonal antibody for the NH2-terminal region of fibrin. Mol Immunol 1984; 21: 89-94
- 5 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
- 6 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
- 7 Feitsma RIJ, Blok D, Wasser MNJM, Nieuwenhuizen W, Pauwels EKJ. A new method for Tc99m-labelling of proteins with an application to clot detection with an antifibrin monoclonal antibody. Nucl Med Commun 1987; 8: 771-777
- 8 Ito RK, Davis GL, Yunis EJ, Houranieh A, Statland BE. Immuno-fibrinolysis: Human fibrin-specific monoclonal antibodies as carriers for fibrinolytic agents. Thromb Haemostas 1985; 54: 276
- 9 Tymkewycz PM, Creighton LJ, Gascoine PS, Zanelli GD, Webbon PM, Gaffney PJ. Imaging of human thrombi in the rabbit jugular vein: I Comparison of two fibrin specific monoclonal antibodies. Thromb Res 1989; 54: 411-421
- 10 Rosebrough SF, Grossman ZD, McAfee JG, Kudryk BJ, Subramanian G, Ritter-Hrncirik CA, Witanowski LS, Tillapaugh-Fay G, Urrutia E, Zapf-Longo C. Thrombus imaging with indium-111 and iodine-131 – labelled fibrin specific monoclonal antibody and its F(ab)2 and Fab fragments. J Nucl Med 1988; 29: 1212-1222
- 11 Hui KY, Haber E, Matsueda GR. Immuno-detection of human fibrin using monoclonal antibody-64C5 in an extracorporeal chick model. Thromb Haemostas 1984; 54: 524-527
- 12 Rosebrough SF, Kudryk B, Grossman ZD, McAfee JG, Subramanian G, Ritter-Hrncirik CA, Witanowski LS, Tillapaugh-Fay G. Radio-immuno imaging of venous thrombi using iodine-131 monoclonal antibody. Radiology 1985; 156: 515-517
- 13 Gaffney PJ, Joe F, Mahmoud M. Giant fibrin fragments derived from crosslinked fibrin. Structure and clinical implication. Thromb Res 1980; 20: 647-662
- 14 Tymkewycz PM, Gaffney PJ. Acquisition of fibrin-specific monoclonal antibodies: Relevance of screening procedures. Fibrinogen III: Biochemistry, Biological Function, Gene Regulation and Expression. Excerpta Medica: International Congress Series No 801 1988: 141-146
- 15 Spitz M, Spitz L, Thorpe R, Eugui E. Intrasplenic primary immunisation for the production of monoclonal antibodies. J Immun Methods 1984; 70: 39-43
- 16 Thorpe R, Perry MJ, Callus M, Gaffney PJ, Spitz M. Single shot intrasplenic immunisation: an advantageous procedure for production of monoclonal antibodies to human fibrin fragments. Hybridoma 1984; 3: 381-385
- 17 Gaffney PJ, Creighton LJ, Perry MJ, Callus M, Thorpe R, Spitz M. Monoclonal antibodies to crosslinked fibrin degradation products (XL-FDP): I. Characterisation and evaluation in plasma. Br J Haematol 1988; 68: 83-90
- 18 Rylatt DB, Blake AS, Cottis LE, Massingham DA, Fletcher WA, Masci PP, Whitaker AN, Elms M, Bunce I, Webber AJ, Wyatt D, Bundesen PG. An immunoassay for human D-dimer using monoclonal antibodies. Thromb Res 1983; 31: 767-778
- 19 Whitaker AN, Elms MJ, Masci PP, Bundesen PG, Rylatt DB, Webber AJ, Bunce IH. Measurement of crosslinked fibrin derivatives in plasma: an immunoassay using monoclonal antibodies. J Clin Pathol 1984; 37: 882-887
- 20 Gaffney PJ, Brasher M. Subunit structure of the plasmin-induced degradation products of crosslinked fibrin. Biochim Biophys Acta 1973; 295: 308-313
- 21 Kudryk B, Rohoza A, Ahadi M, Chin J, Wiebe ME. A monoclonal antibody with ability to distinguish between NH2-terminal fragments from fibrinogen and fibrin. Mol Immunol 1983; 20: 1191-1200
- 22 Kudryk B, Grossman ZD, McAfee JG, Rosebrough SF. Monoclonal antibodies as probes for fibrin(ogen) proteolysis. In: Monoclonal Antibodies in Immunoscintigraphy. CRC Press; Boca Raton, FL: 1989: 365-398
- 23 Tymkewycz PM, Gascoine PS, Gaffney PJ. A monoclonal antibody (MAB) to an alpha chain epitope exposed only on the surface of fibrin. Thromb Haemostas 1989; 62: 478
- 24 Gaffney PJ, Gascoine PS, Creighton LJ, Tymkewycz PM. Monoclonal antibodies for the detection of thrombosis. Adv Exp Med Biol 1990; 281: 419-427
- 25 Gargan PE, Ploplis VA, Pollard M, Scheu J. A fibrin specific monoclonal antibody which interferes with tissue plasminogen activator binding to fibrin. Proc VIIIth Int Congr Fibrinolysis. 1986 (Abstr 128)
- 26 Hockley DJ, Wood RD, Jacobs JP, Garrett AJ. Electron microscopy of human immunodeficiency virus. J Gen Virol 1988; 69: 2455-2469
- 27 Soria J, Soria C, Mirshahi M, Boucheix C, Aurengo A, Perrot J, Bernadou A, Samama M, Rosenfeld C. Conformational change in fibrinogen induced by adsorption to a surface. J Colloid Interface Sci 1985; 107 (1) 204-208
- 28 Morrissey BW. The adsorption and conformation of plasma proteins: A physical approach. Ann NY Acad Sci 1977; 283: 50-64
- 29 Chuang MYK, King WF, Mason RG. Interaction of plasma proteins with artificial surfaces: Protein adsorption isotherms. J Lab Clin Med 1978; 92: 483-496
- 30 Gaffney PJ, Creighton-Kempsford LJ, Tymkewycz PM. Characterisation profile of five monoclonal antibodies with an affinity for fibrin. Thromb Haemostas 1991; 65: 903