Abstract
The product of the c-erbB-2 protooncogene (p185) is a member of the EGF receptor family of transmembrane tyrosine kinases. Amplification of this gene and overexpression of the product has been observed in adenocarcinomas and has been correlated with poor prognosis in patients with breast and ovarian cancer. The very low levels of expression of p185 by normal adult tissues makes the receptor an almost tumor-specific target. We have prepared rat monoclonal antibodies against five distinct epitopes on the external domain of the c-erbB-2 product overexpressed by the breast cancer line BT474. The antibodies bind to the protein core of p185 and stain specifically the membranes in frozen sections of tumors overexpressing the c-erbB-2 product. Three of the antibodies, ICR12 (epitope A), ICR54, and ICR55 (epitope E), also stain the cell membrane in formalin-fixed, paraffin-embedded sections and bind to p 185 in Western blots. An investigation of the stability of the antigen-antibody complexes indicates that the majority are not readily internalized by SKOV3 cells growing in vitro. Antibodies ICR12 (IgG2a) and ICR55 (IgG2a), which are directed against separate epitopes on the c-erbB-2 p185, are both of high affinity and immunoreactivity (>75%) and localize specifically and stably to xenografted breast and ovarian carcinomas growing in athymic mice when labeled with125I (up to 13% injected dose/g, ICR12 and ICR55) or a range of other radionuclides (up to 20% id/g, ICR12). We conclude that these antibodies may be useful as therapeutic vehicles for targeting radionuclides (imaging and therapy) or enzymes for activating prodrugs (ADEPT).
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Kohler, G. and Milstein, C. (1975) Continuous culture of fused cells secreting antibody of predefined specificity.Nature 256, 495–497.
Gullick, W. J. (1991) Prevalence of aberrant expression of the epidermal growth factor receptor in human cancers.Br. Med. Bull. 47(1), 87–98.
Masui, H. Kawamoto, T., Sato, J. D., Wolf, B., Sato, G., and Mendelsohn, J. (1984) Growth inhibition of human tumor cells in athymic mice by anti-epidermal growth factor receptor monoclonal antibodies.Cancer Res.44, 1002–1007.
Rodeck, U., Herlyn, M., Herlyn, D. Molthoff, C., Atkinson, B., Varello, M., Steplewski, Z., and Koprowski, H. (1987) Tumor growth modulation by a monoclonal antibody to the epidermal growth factor receptor: immunologically mediated and effector cell-independent effects.Cancer Res. 47, 3692–3696.
Abound-Pirak, E., Hurwitz, E., Pirak, M. E., Bellot, F., Schlessinger, J., and Sela, M. (1988) Efficacy of antibodies to epidermal growth factor receptor against KB carcinoma in vitro and in nude mice.J. Natl. Cancer Inst. 80, 1605–1611.
Akiyama, T., Sudo, C. Ogawara, M., Toyoshima, K., and Yamamoto, T. (1986) The product of the c-erbB-2 gene: a 185-kilodalton glycoprotein with tyrosine kinase activityScience 232, 1644–1646.
Gullick, W. J., Berger, M.S., Bennett, P. L. P., Rothbard, J. G., and Waterfield, M. D. (1987) Expression of the c-erbB-2 protein in normal and transformed cells.Int. J. Cancer 40, 246–254.
Slamon, D. J., Clark, G. M., Wong, S. G., Levin, W. J., Ullrich, A., and McGuire, W. L. (1987) Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene.Science 235, 177–182.
Gusterson, B. A., Machin, L. G., Gullick, W. J., Gibbs, N. M., Powles, T. J., Elliott, C., Ashley, S., Monaghan, P., and Harrison, S. (1988) c-erbB-2 expression in benign and malignant disease.Br. J. Cancer 58, 453–457.
Gusterson, B. A., Gelber, R. D., Goldhirsch, A., Price, K. N. Säve-Söderborgh, J., Anbazhagan, R., Syles, J., Rudenstam, C.-M., Golough, R., Reed, R., Matinez-Tello, F., Titman, A., Torhorst, J., Grigolato, P., Bettelhein, R., Neville, A. M., Bürki, K., Castiglione, M., Collins, J., Lindtner, J., and Senn, H.-J. (1992) Prognostic importance of c-erbB-2 expression in breast cancer.J. Clin. Oncol. 10, 1049–1056.
De Potter, C. R., Beghin, C., Makar, A. P., Vandeerckhove, D., and Roels, J. (1990) Theneu oncogene protein as a predictive factor for haematogenous metastases in breast cancer patients.Int. J. Cancer 45, 55–58.
Ro, J. El-Naggar, A., Ro, J. Y., Blick, M., Fraschini, F., Fritsche, H., and Hortobagyi, G. (1989) c-erbB-2 amplification in node negative breast cancer: a positive relation with nuclear aplasia and DNA aneuploidy. A possible prognostic factor.Lab. Invest. 60, 70A.
Styles, J. M., Harrison, S., Gusterson, B. A., and Dean, C. J. (1990) Rat monoclonal antibodies to the external domain of the product of the c-erbB-2 proto-oncogene.Int. J. Cancer 45, 320–324.
Dean, C., Styles, J., Valeri, M., Modjtahedi, H., Bakir, A., Babich, J., and Eccles, S. (1993) Growth factor receptors as targets for antibody therapy, inMutant Oncogenes: Targets for Therapy? (Epenetos, A. A. and Lemoine, N. R., eds.), Chapman and Hall, New York, London, pp. 27–34.
Allan, S., Dean, C., Fernando, I., Eccles, S., Styles, J., McCready, V. R. Baum, M., and Sacks, N. (1993) Radioimmunolocalisation in breast cancer using the gene product of the c-erbB-2 as target antigen.Br. J. Cancer 67, 706–712.
Dean, C. J., Styles, J. M., Gyure, L. A., Peppard, J., Hobbs, S. M., Jackson, E., and Hall, J. G. (1984) The production of hybridomas from the gut associated lymphoid tissue of tumor bearing rats. I. Mesenteric nodes as a source of IgG producing cells.Clin. Exp. Immunol. 57, 358–364.
Fraker, P. J. and Speck, J. C. (1978) Protein and cell membrane iodination with sparingly soluble chloramide 1, 3, 4, 4-tetra chloro-3-6-diphenylglycouril.Biochem. Biophys. Res. Comm. 80, 849.
Bakir, M. A., Eccles, S. A., Babich, J. W., Aftab, N., Styles, J. M., Dean, C. J., and Ott, R. W. (1992) C-erb B-2 protein overexpression in breast cancer as a target for P.E.T. using1-I labelled monoclonal antibodies.J. Nucl. Med. 33, 2154–2160.
Krejcarek, G. E. and Tucker, K. L. (1977) Covalent attachment of chelating groups to macromolecules.Biochem. Biophys. Res. Comm. 77, 581.
Saga, T., Endo, K., Akiyama, T., Sakahara, H., Koizumi, M., Watanabe, Y., Nakai, T., Hosono, M., Yamamoto, T., Toyoshima, K., and Konishi, J. (1991) Scintigraphic detection of overexpressed c-erbB-2 protooncogene products by a class-switched murine anti-c-erbB-2 protein monoclonal antibody.Cancer Res. 51, 990–994.
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Dean, C.J., Eccles, S.A., Valeri, M. et al. Rat MAbs to the product of the c-erbB-2 proto-oncogene for diagnosis and therapy in breast cancer. Cell Biophysics 22, 111–127 (1993). https://doi.org/10.1007/BF03033870
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DOI: https://doi.org/10.1007/BF03033870