Summary
A microassay carried out with V-bottomed microplates was developed as a simplified procedure to estimate the complement hemolytic activity. This microassay has the advantage of permitting the evaluation of the extent of lysis without the need to separate the supernatants from the unlysed erythrocytes. This is accomplished by calculating the ratio between the absorbance at 405 nm and that at 450 nm. The results, read automatically on a Dynatech MR580 spectrophotometer, are transformed into CH50 U by means of a computer program. Finally, a kinetic assessment of complement hemolytic activity, easily carried out by this simple procedure, may reveal complement abnormalities otherwise undetected by the standard hemolytic assay.
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References
Bliss C. I.: Statistics in biology. McGraw-Hill, New York, 1970.
Finney D. J.: Probit analysis. Cambridge University Press, Cambridge, 1971.
Gewurz H., Suyehira L. A.: Complement. In:Rose N. R., Friedman H. (Eds): Manual of clinical immunology. American Society for Microbiology, Bethesda, 1976; p. 36.
Klerx J. P. A. M., Beukelman C. J., Van Dijk H., Willers J. M. N.: MIcroassay for colorimetric estimation of complement activity in guinea pig, human and mouse serum — J. immunol. Meth.63, 215, 1983.
Lachmann P. J., Hobart M. J.: Complement technology. In:Weir D. M. (Ed.): Handbook of experimental immunology. Blackwell Sci. Publ., Oxford, 1978; chapt. 5A.
Mayer M. M.: Complement and complement fixation. In:Kabat E. A., Mayer M. M. (Eds): Experimental immunochemistry. 2nd ed., C. C. Thomas Publ., Springfield, Ill., 1971; p. 133.
Nilsson V. R., Nilsson B.: Simplified assays of haemolytic activity of the classical and alternative complement pathways — J. immunol. Meth.72, 49, 1984.
Notarangelo L. D., Plebani A., Marconi M., Chiara A., Martini A., Ugazio A. G.: A simple hemolytic micromethod for the quantitative and kinetic evaluation of the alternative activating pathway of complement — La Ricerca Clin. Lab.13, 423, 1983.
Pearson E. S., Hartley H. O.: Biometrical tables for statisticians. Cambridge University Press, Cambridge, 1954.
Platts-Mills T. A. E., Ishizaka K.: Activation of the alternative pathway of human complement by rabbit cells — J. Immunol.113, 348, 1974.
Salkie M. L., Kelker D. H.: An investigation into the relationship between serum levels of C3, C4 and CH50 — Vox Sang. (Basel)41, 282, 1981.
Sims P. J., Boswell E. B.: The measurement of platelet-associated IgG by a microplate quantitative antiglobulin consumption assay using automated through-the-well spectrophotometry — J. Lab. clin. Med.102, 352, 1983.
Takada A., Imamura Y., Takada Y.: Relationship between the haemolytic activities of the human complement system and complement components — Clin. exp. Immunol.35, 324, 1979.
Tedesco F., Bardare S. M., Giovanetti A. M., Sirchia G.: A familial dysfunction of the eighth component of complement (C8) — Clin. Immunol. Immunopathol.16, 180, 1980.
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This work was supported by a grant from theConsiglio Nazionale delle Ricerche (CNR), Roma, Italy,Progetto Finalizzato ‘Ingegneria Genetica e Basi Molecolari delle Malattie Ereditarie’.
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Narchi, G., Milani, S. & Tedesco, F. A computerized microassay for complement hemolytic activity using an automated spectrophotometer. La Ricerca Clin. Lab. 17, 27–33 (1987). https://doi.org/10.1007/BF02909385
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DOI: https://doi.org/10.1007/BF02909385