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

Russian Journal of Bioorganic Chemistry volume 26, pages 482–489 (2000)Cite this article

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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Authors and Affiliations

  1. Gabrichevskii Moscow Research Institute of Epidemiology and Microbiology, ul. Admirala Makarova 10, 125212, Moscow, Russia

    L. V. Kozlov, V. M. Lakhtin, T. G. Skorokhodova, T. N. Batalova, B. B. Shoibonov, V. L. D’yakov, V. A. Guzova & N. S. Matveevskaya

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  1. L. V. Kozlov
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  2. V. M. Lakhtin
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  3. T. G. Skorokhodova
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  4. T. N. Batalova
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  5. B. B. Shoibonov
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  6. V. L. D’yakov
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  7. V. A. Guzova
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  8. N. S. Matveevskaya
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Correspondence to L. V. Kozlov.

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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