Abstract
The difference in the functional activity of the isotypes A and B of component C4 of human complement was used to determine their ratio and to detect the inherited deficiency of the isotypes. ELISA methods were developed for the quantitative assay of component C4 (conventional sandwich method) and its functional activity. When determining the functional activity, the classic pathway of the complement and therefore of component C4 was activated on activators sorbed on ELISA microplates: immunoglobulin IgG3 or liposaccharide of theShigella sonnei cell walls, which activates the complement by binding component C1. The nascent fragment C4b is covalently bound to the target activator; C4Ab binds better to the target protein (immunoglobulin), and C4Bb to the target carbohydrate (liposaccharide). Therefore, when immunoglobulin is a target activator, isotype C4A is bound and determined; and when the complement is activated with liposaccharide, isotype C4B is determined. The radio of the activities determined by the two methods indicates the deficiency in the individual isotypes of component C4 or its absence. The rabbit polyclonal monospecific antibodies against the human component C4 and the conjugates of these antibodies with horseradish peroxidase were used in the methods described.
Similar content being viewed by others
References
Low, S.K.A. and Dodds, A.W.,Protein Science, 1997, vol. 6, pp. 265–274.
Manuff, G., Alper, C.A., Awdeh, Z., Batchelor, R., Bertrams, J., Bruun-Petersen, G., Dawkins, R.L., Démant, P., Edwards, J., Grosse-Wilde, H., Hauptmann, G., Klouda, P., Lamm, L., Mollenhauer, E., Nerl, C., Olaisen, B., O’Neill, G., Rittner, C., Roos, M.H., Skanes, V., Teisberg, P., and Wells, L.,Immunobiology, 1983, vol. 164, pp. 184–191.
O’Neill, G.J., Yang, S.Y., Tegoli, J., Berger, R., and Dupont, B.,Nature, 1978, vol. 273, pp. 668–670.
Reilly, B.D. and Mold, C.,Clin. Exp. Immunol., 1997, vol. 110, pp. 310–316.
Brai, M., Accardo, P., and Bellavia, D.,Ann. Ital. Med. Int., 1994, vol. 9, pp. 167–172.
Fielder, A.H., Walport, M.J., Batchelor, J.R., Rynes, R.I., Black, C.M., Dodi, I.A., and Hughes, G.R.,Br. Med. J., 1983, vol. 286, pp. 425–428.
Bertrams, J., Hintzen, U., Schlicht, V., Schoeps, S., Gries, F.A., Louton, T.K., and Baur, M.P.,Immunobiology, 1984, vol. 166, pp. 335–344.
Vergani, D., Wells, L., Lacher, V.F., Nasaruddin, B.A., Davies, E.T., Mieli-Vergani, G., and Mowat, A.P.,Lancet, 1985, vol. 2, pp. 294–298.
Kramer, J., Rajczy, K., and Füst, G.,Immunol. Lett., 1989, vol. 20, pp. 83–86.
Law, S.K.A., Dodds, A.W., and Porter, R.R.,EMBO J., 1984, vol. 3, pp. 1819–1823.
Kozlov, L.V., Shibanova, E.D., and Zinchenko, A.A.,Biokhimiya, 1987, vol. 52, pp. 660–666.
Kozlov, L.V., Batalova, T.N., Faddeeva, T.V., and Panurina, R.L.,Bioorg. Khim., 1997, vol. 23, pp. 635–641.
Kozlov, L.V., Krylova, Yu.I., Chikh, V.P., and Molchanova, N.N.,Bioorg. Khim., 1982, vol. 8, pp. 652–659.
Kozlov, L.V., Sizoi, M.N., Zinchenko, A.A., Ivanov, A.E., and Zubov, V.P.,Bioorg. Khim., 1984, vol. 10, pp. 1629–1638.
Kozlov, L.V., Zinchenko, A.A., Solyakov, L.S., Sizoi, M.N., Ishchenko, A.M., Martyushin, S.V., and Andreev, S.V.,Bioorg. Khim., 1983, vol. 9, pp. 1047–1055.
Morrison, D.C. and Kline, L.F.,J. Immunol., 1977, vol. 118, pp. 362–368.
Sepp, A., Dodds, A.W., Anderson, M.J., Campbell, R.D., Willis, A.C., and Law, S.K.,Protein Science, 1993, vol. 2, pp. 706–716.
O’Neill, G.J., Yang, S.Y., and Dupont, B.,Proc. Natl. Acad. Sci. USA, 1978, vol. 75, pp. 5165–5169.
Awdeh, Z.L., Raum, D., and Alper, C.A.,Nature, 1979, vol. 282, pp. 205–207.
Skorokhodova, T.G., Kozlov, L.V., Batalova, T.N., and Lakhtin, V.M.,Klinicheskaya laboratornaya diagnostika, 1999, no. 9, p. 34.
Getty, D., Raykundalia, C., and Houba, V.,WHO IMM/PIR, 1983, no. 83.1.
Westphal, O. and Jann, K.,Methods in Carbohydrate Chemistry, Wistler, R.L., Ed., New York: Academic, 1965, vol. 5, pp. 83–91.
Kozlov, L.V., D’yakov, V.L., Mishin, A.A., Guzova, V.A., Batalova, T.N., and Lysakova, S.V, RF Patent 2144365,Byull. Izobret., 2000, no. 2.
Ouchterlony, O. and Nilsson, L.-A.,Handbook of Experimental Immunology, Oxford: Blackwells, 1978, vol. 1, pp. 19.1–19.44.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kozlov, L.V., Lakhtin, V.M., Skorokhodova, T.G. et al. Isotyping of component C4 of human complement using differences in the functional activity of isotypes C4A and C4B. Russ J Bioorg Chem 26, 482–489 (2000). https://doi.org/10.1007/BF02758619
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF02758619