Abstract
We describe the production and characterization of a monoclonal antibody specific for platelets. This antibody reacts strongly with human and primate platelets, but does not recognise human monocytes, polymorphonuclear leucocytes, lymphocytes, erythrocytes, leukaemic nor fibroblast cell lines, nor rodent platelets. Immunoprecipitation studies using radiolabelled platelet membrane proteins showed that the monoclonal antibody binds to the platelet membrane glycoprotein IIb-IIIa complex. Affinity chromatography using immobilized monoclonal antibody allows purification of the antigen, but also co-purifies the cytoskeletal proteins actin and myosin.
Our results demonstrate immunochemically that although the GP IIb-IIIa complex is an external structure, it is connected through the cell membrane to the microfilament system.
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References
Bai, Y., Durbin, H., and Hogg, N. (1984). Monoclonal antibodies specific for platelet glycoproteins react with human monocytes.Blood 64(1):139–146.
Berndt, M. C., and Phillips, D. R. (1981). Platelet membrane proteins: composition and receptor function. InPlatelets in Biology and Pathology-2 (J. L. Gordon Ed.), Elsevier/North Holland Biomedical Press.
Galfre, G., Howe, S. C., Milstein, C., Butcher, G. W., and Howard, J. C. (1977). Antibodies to major histocompatibility antigens produced by hybrid cell lines.Nature 266:550–552.
Gearing, A. J. H., Thorpe, R., Spitz, L., and Spitz, M. (1984). Use of “single shot” intrasplenic immunization for production of monoclonal antibodies specific for human IgM..Journal of Immunological Methods 76:337–343.
Graham, R. C., and Karnovsky, M. J. (1966). The early stages of absorption of horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique.J. Histochem. Cytochem. 14:291–302.
Higashihara, M., Maeda, H., Shibata, Y., Kume, S., and Ohashi, T. (1985). A monoclonal anti-human platelet antibody: a new platelet aggregating substance.Blood 65(2):382–391.
Johnstone, A., and Thorpe, R. (1982).Immunochemistry in Practice, Blackwell Scientific Publications.
Jones, D., Fritschy, J., Garson, J., Nokes, T. J. C., Kemshead, J. T., and Hardisty, R. M., (1984). A monoclonal antibody binding to human medulloblastoma cells and to the platelet glycoprotein IIb-IIIa complex.British Journal of Haematology 57:621–631.
Laemmli, U. K., and Favre, M. J. (1973). Maturation of the head bacteriophage T4.Journal of Molecular Biology 80:575–590.
Menashi, S., Weintroub, H., and Crawford, N. (1981). Characterization of human platelet surface and intracellular membranes isolated by free flow electrophoresis.Journal of Biological Chemistry 256:4095–4101.
Phillips, D. R., Jennings, L. K., and Edwards, H. H. (1980). Identification of membrane proteins mediating interaction of human platelets.Journal of Cell Biology 86:77–86.
Reichlin, M. (1975). Amino acid substitution and the antigenicity of globular proteins.Advances in Immunology 20:71–123.
Rotman, A., Heldman, J., and Linder, S. (1982). Association of membrane and cytoplasmic proteins with the cytoskeleton in blood platelets.Biochemistry 21:1713–1719.
Sela, M. (1969). Antigenicity: some molecular aspects.Science 166:1365–1374.
Thorpe, R., Bird, C. R., and Spitz, M. (1984). Immunoblotting with monoclonal antibodies: loss of immunoreactivity with human immunoglobulins arises from polypeptide chain separation.Journal of Immunological Methods 73:259–265.
Wray, W., Boulikas, T., Wray, V. P., and Hancock, R. (1981). Silver staining of proteins in polyacrylamide gels.Analytical Biochemistry 118:197–203.
Zola, H., McNamara, P. J., Beckman, I. G. R., Brooks, D. A., Macardle, P. J., Bradley, J., Goodall, K., Skinner, J., Collins, R., and Berndt, M. C. (1984). Monoclonal antibodies against antigens of the human platelet surface: preparation and properties.Pathology 16:73–78.
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Bird, C., Callus, M., Trickett, L. et al. Immunochemical characterization of a new platelet specific monoclonal antibody and its use to demonstrate the cytoskeletal association of the platelet glycoprotein IIb-IIIa complex. Biosci Rep 6, 323–333 (1986). https://doi.org/10.1007/BF01115162
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DOI: https://doi.org/10.1007/BF01115162