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Isotyping of component C4 of human complement using differences in the functional activity of isotypes C4A and C4B

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.

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References

  1. Low, S.K.A. and Dodds, A.W.,Protein Science, 1997, vol. 6, pp. 265–274.

    Google Scholar 

  2. 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.

    Google Scholar 

  3. O’Neill, G.J., Yang, S.Y., Tegoli, J., Berger, R., and Dupont, B.,Nature, 1978, vol. 273, pp. 668–670.

    PubMed  Article  CAS  Google Scholar 

  4. Reilly, B.D. and Mold, C.,Clin. Exp. Immunol., 1997, vol. 110, pp. 310–316.

    PubMed  CAS  Article  Google Scholar 

  5. Brai, M., Accardo, P., and Bellavia, D.,Ann. Ital. Med. Int., 1994, vol. 9, pp. 167–172.

    PubMed  CAS  Google Scholar 

  6. 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.

    CAS  Google Scholar 

  7. 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.

    PubMed  CAS  Google Scholar 

  8. 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.

    PubMed  Article  CAS  Google Scholar 

  9. Kramer, J., Rajczy, K., and Füst, G.,Immunol. Lett., 1989, vol. 20, pp. 83–86.

    PubMed  Article  CAS  Google Scholar 

  10. Law, S.K.A., Dodds, A.W., and Porter, R.R.,EMBO J., 1984, vol. 3, pp. 1819–1823.

    PubMed  CAS  Google Scholar 

  11. Kozlov, L.V., Shibanova, E.D., and Zinchenko, A.A.,Biokhimiya, 1987, vol. 52, pp. 660–666.

    CAS  Google Scholar 

  12. Kozlov, L.V., Batalova, T.N., Faddeeva, T.V., and Panurina, R.L.,Bioorg. Khim., 1997, vol. 23, pp. 635–641.

    PubMed  CAS  Google Scholar 

  13. Kozlov, L.V., Krylova, Yu.I., Chikh, V.P., and Molchanova, N.N.,Bioorg. Khim., 1982, vol. 8, pp. 652–659.

    CAS  Google Scholar 

  14. Kozlov, L.V., Sizoi, M.N., Zinchenko, A.A., Ivanov, A.E., and Zubov, V.P.,Bioorg. Khim., 1984, vol. 10, pp. 1629–1638.

    PubMed  CAS  Google Scholar 

  15. 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.

    PubMed  CAS  Google Scholar 

  16. Morrison, D.C. and Kline, L.F.,J. Immunol., 1977, vol. 118, pp. 362–368.

    PubMed  CAS  Google Scholar 

  17. 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.

    PubMed  CAS  Article  Google Scholar 

  18. O’Neill, G.J., Yang, S.Y., and Dupont, B.,Proc. Natl. Acad. Sci. USA, 1978, vol. 75, pp. 5165–5169.

    PubMed  Article  CAS  Google Scholar 

  19. Awdeh, Z.L., Raum, D., and Alper, C.A.,Nature, 1979, vol. 282, pp. 205–207.

    PubMed  Article  CAS  Google Scholar 

  20. Skorokhodova, T.G., Kozlov, L.V., Batalova, T.N., and Lakhtin, V.M.,Klinicheskaya laboratornaya diagnostika, 1999, no. 9, p. 34.

  21. Getty, D., Raykundalia, C., and Houba, V.,WHO IMM/PIR, 1983, no. 83.1.

  22. Westphal, O. and Jann, K.,Methods in Carbohydrate Chemistry, Wistler, R.L., Ed., New York: Academic, 1965, vol. 5, pp. 83–91.

    Google Scholar 

  23. 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.

  24. Ouchterlony, O. and Nilsson, L.-A.,Handbook of Experimental Immunology, Oxford: Blackwells, 1978, vol. 1, pp. 19.1–19.44.

    Google Scholar 

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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

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  • DOI: https://doi.org/10.1007/BF02758619

Key words

  • complement system
  • isotypes C4A and C4B
  • functional activity assay
  • ELISA