Abstract
Cross-reactions among carbonic anhydrases (CAs) I, II, and III were studied using a variety of antisera: (1) a rabbit antiserum to bovine CA III, (2) mouse antisera to human CA I, CA II, and CA III; and (3) five monoclonal antibodies prepared by the hybridoma technique using splenocytes from a mouse immunized with human CAs I and II and bovine CA III. Cross-reactions between CAs were readily found by binding assays using these antisera. Human CA I, but not human CA II, inhibited the reaction of the rabbit anti-CA III with its homologous antigen. Mouse antisera to CA I or CA II bound the homologous I or II with nearly as great efficiency as the autologous isozyme and sometimes weakly bound CA III. Mouse antisera to CA III frequently bound CA I or II. These cross-reactions were confirmed by the first use of hybridoma-prepared, monoclonal antibodies to CAs. The mouse monoclonal antibodies to CA isozymes varied in the amount of cross-reactivity among I, II, and III: at one extreme, one monoclonal was highly specific for the autologous CA III; at the other extreme, one monoclonal weakly reacted with some examples of CAs I, II, and III.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Arnon, R., Sela, M., Yaron, A., and Sober, H. A. (1965). Polylysine-specific antibodies and their reaction with oligolysines. Biochemistry 4948.
Carter, N. D., Hewett-Emmett, D., Jeffery, S., and Tashian, R. E. (1981). Testosterone-induced carbonic anhydrase isozyme of rat liver is indistinguishable from skeletal muscle carbonic anhydrase III. FEBS Lett. 128114.
Claflin, J. L., Hudak, S., and Maddalena, A. (1981). Anti-phosphorylcholine hybridoma antibodies. I. Direct evidence for three distinct families of antibodies in the murine response. J. Exp. Med. 153352.
Dodgson, S. J., Forster, R. E., Storey, B. T., and Mela, L. (1980). Mitochondrial carbonic anhydrase. Proc. Natl. Acad. Sci. USA 775562.
Dulbecco, R., Unger, M., Bologna, M., Battifora, H., Syka, P., and Okada, S. (1981). Cross-reactivity between Thy-1 and a component of intermediate filaments demonstrated using a monoclonal antibody. Nature 292772.
Erickson, R. P., and Rosenberg, L. T. (1973). The role of tolerance in the lack of cross-reactivity between homologous (structurally related) proteins: Cross-reactions between α-lactalbumins and between lysozymes recognized by antibodies from intolerant rabbits. J. Immunochem. 10467.
Fine, J. M., and Laurent, G. (1970). Immunological studies of the carbonic anhydrases isolated from human and animal erythrocyte hemolysates. Progr. Immunobiol. Stand. 4411.
Fraker, P. J., and Speck, J. C., Jr. (1978). Protein and cell membrane iodinations with a sparingly soluble chloramide 1, 3, 4, 6-tetrachloro-3α, 6α-diphenyl-glycoluril. Biochem. Biophys. Res. Comm. 80849.
Gefter, M. L., Margulies, D. H., and Scharff, M. D. (1977). A simple method for polyethylene glycol-promoted hybridization of mouse myeloma cells. Somat. Cell Genet. 3231.
Herzenberg, L. A., and Herzenberg, L. A. (1978). Mouse immunoglobulin allotypes. In Weir, D. M. (ed.), Handbook in Experimental Immunology 3rd ed., Blackwell Scientific, Oxford, p. 12.21.
Kabat, E. A. (1966). The nature of an antigenic determinant. J. Immunol. 971.
Kannan, K. K. (1980). Crystal structure of carbonic anhydrase. In Baurer, C., Gros, G., and Bartels, H. (eds.), Biophysics and Physiology of Carbon Dioxide Springer-Verlag, New York, pp. 184–205.
Kishida, K., and Ochi, N. (1980). Evidence for the existence of two carbonic anhydrase isozymes in rabbit ciliary body. Experientia 3642.
Klinman, N. R. (1972). The mechanism of antigenic stimulation of primary and secondary clonal precursor cells. J. Exp. Med. 136241.
Landsteiner, K. (1946). The Specificity of Serological Reactions rev. ed., Dover reprint of 1962, Dover, New York, pp. 270–272.
Littlefield, J. W. (1964). Selection of hybrids from matings of fibroblasts in vitro and their presumed recombinants. Science 145709.
Mandal, C., and Karush, F. (1981). Restriction in IgM expression. III. Affinity analysis of monoclonal anti-lactose antibodies. J. Immunol. 1271240.
Margulies, D. H., Kuehl, W. M., and Scarff, M. D. (1976). Somatic cell hybridization of mouse myeloma cells. Cell 8405.
McKinley, D. N., and Whitney, P. L. (1976). Particulate carbonic anhydrase in homogenates of human kidneys. Biochim. Biophys. Acta 445780.
Michili, A., and Buzzi, C. (1964). Electrophoretic and immuno-electrophoretic studies on the carbonic anhydrase of the human red blood cells. Biochim. Biophys. Acta 89324.
Nonno, L., Herschman, H., and Levine, L. (1969). Serologic comparisons of the carbonic anhydrases of primate erythrocytes. Arch. Biochem. Biophys. 136361.
Osborne, W. R. A., and Tashian, R. E. (1975). An improved method for the purification of carbonic anhydrase isozymes by affinity chromatography. Anal. Biochem. 64297.
Pillemer, E., and Weissman, I. L. (1981). A monoclonal antibody that detects a V-TEPC15 idiotypic determinant cross-reactive with a Thy-1 determinant. J. Exp. Med. 1531068.
Pressman, D., and Grossberg, A. L. (1973). The Structural Basis of Antibody Specificity W. A. Benjamin, Reading, Pa., pp. 148–165.
Rudikoff, S., Potter, M., Segal, D. M., Padlan, E. A., and Davies, D. R. (1972). Crystals of phosphorylcholine-binding Fab-fragments from mouse myeloma proteins: Preparation and X-ray analysis. Proc. Natl. Acad. Sci. USA 693689.
Suyama, H., Sawada, H., Ueda, H., and Ohya, I. (1968a). Erythrocyte carbonic anhydrases of the human and some animals. 3. Immunological study of human erythrocyte carbonic anhydrases. Med. Biol. (Jap.) 7715.
Suyama, H., Sawada, H., Ueda, H., and Ohya, I. (1968b). Erythrocyte carbonic anhydrases of the human and some animals. 4. Immunological study of animal erythrocyte carbonic anhydrases. Med. Biol. (Jap.) 7721.
Tashian, R. E. (1977). Evolution and regulation of the carbonic anhydrase isozymes. In Rattazzi, M. C., Scandalios, J. G., and Whitt, G. S. (eds.), Isozymes: Current Topics in Biological and Medical Research, Vol. 2 Alan R. Liss, New York, pp. 21–62.
Tashian, R. E., Shreffler, D. C., and Shows, T. B. (1968). Genetic and phylogenetic variation in the different molecular forms of mammalian erythrocyte carbonic anhydrases. Ann. N.Y. Acad. Sci. 15164.
Tashian, R. E., Hewett-Emmett, D., and Goodman, M. (1980a). Evolutionary diversity in the structure and activity of carbonic anhydrase. In Peeters, H. (ed.), Protides of the Biological Fluids, Vol. 28 Pergamon Press, Oxford, pp. 153–156.
Tashian, R. E., Hewitt-Emmett, D., Stroup, S. K., Goodman, M., and Yu, Y.-S. L. (1980b). Evolution of structure and function in the carbonic anhydrase isozymes of mammals. In Bauer, C., Gros, G., and Bartels, H. (eds.), Biophysics and Physiology of Carbon Dioxide Springer-Verlag, New York, pp. 165–176.
Twining, S. S., Lehmann, H., and Atassi, M. Z. (1980). The antibody response to myoglobin is independent of the immunized species. Biochem. J. 191681.
Author information
Authors and Affiliations
Additional information
This work was supported by NIH Grant GM-24681 and a grant from the National Foundation-March of Dimes.
Rights and permissions
About this article
Cite this article
Erickson, R.P., Kay, G., Hewett-Emmett, D. et al. Cross-reactions among carbonic anhydrases I, II, and III studied by binding tests and with monoclonal antibodies. Biochem Genet 20, 809–819 (1982). https://doi.org/10.1007/BF00483975
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00483975