THE DEVELOPMENT OF TOTAL COMPLEMENT ACTIVITY ASSAY BASED ON COMPLEMENT-DEPENDENT IMMUNE LIPOSOME LYSIS

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Abstract


Background: The purpose of work was development of a fast and reproduced procedure for measurement of the total complement activity (ТСА) in human or animal blood serum. Materials and methods: Steady at storage liposomes preparations, which surface sensitized 2,4-DNP haptens, and the internal volume contains calceine or sulforhodamine 101 are obtained. Complement-dependent immune lysis of liposomes at presence of the anti-2,4-DNP immunoglobulines and complement preparations from animals are investigated. Results: It is shown that the degree of liposomes immune lysis depends on complement concentration in a wide range that can be used for definition of TCA level. Research of blood sera from patients has revealed correlation (r =0,793) between data received with the help of liposome immunolytic systems, and the data of nephelometric analysis with application of suspension sheep erythrocytes. Conclusion: The method allows to define total complement activity in blood serum in 15 minutes without separation of reaction components. This might be useful for measurement ТСА level at patients with various diseases and realization of scientific researches.


S. N. Skopinskaya

State Scientific Research Institute of Biological Engineering, Moscow, Russian Fеderation

Author for correspondence.
Email: diasol@dol.ru

Russian Federation

кандидат биологических наук, ведущий научный сотрудник отдела спектральных методов анализа ГосНИИ биологического приборостроения
Адрес: 125424, Москва, Волоколамское ш., д. 75, тел.: (495) 491-86-83

S. P. Yarkov

State Scientific Research Institute of Biological Engineering, Moscow, Russian Fеderation

Email: diasol@dol.ru

Russian Federation

доктор биологических наук, начальник отдела спектральных методов анализа ГосНИИ биологического приборостроения
Адрес: 125424, Москва, Волоколамское ш., д. 75, тел.: (495) 491-86-83

E. N. Khramov

State Scientific Research Institute of Biological Engineering, Moscow, Russian Fеderation

Email: niibp@dol.ru

Russian Federation доктор технических наук, профессор, директор ГосНИИ биологического приборостроения
Адрес: 125424, Москва, Волоколамское ш., д. 75, тел.: (495) 491-73-72

A. V. Antashev

Academician V.I. Shumakov Federal Research Center of Transplantology and Artificial Organs, Moscow, Russian Fеderation

Email: artem72antashev@mail.ru

Russian Federation

кандидат медицинских наук, врач-лаборант ФНЦ трансплантологии и искусственных органов им. акад. В.И. Шумакова, врач-лаборант Клинической больницы № 1 Управления делами Президента РФ Адрес: 123182, Москва, ул. Щукинская, д. 1, тел.: (495) 491-35-05

  1. Валпорт М. Иммунология. Пер. с англ. Под. ред. А. Ройта, Дж. Бростоффа, Д. Мейла. М.: Мир. 2000. С. 59–81.
  2. Figueroa J.A., Densen P. Infection diseases associated with complement deficiencies. Clin. Microbiol. Rev. 1991; 4: 359–395.
  3. Morgan B.P. Physiology and pathophysiology of complement: progress and trends. Crit. Rev. Clin. Lab. Sci. 1995; 32: 265–298.
  4. Rogers J., Cooper N.R., Webster S. et al. Complement activation by β-amyloid in Alzheimer disease. Proc. Natl. Acad. Sci. USA. 1992; 89: 10016–10020.
  5. Klein M.A., Kaeser P.S., Schwarz P. et al. Complement facilitates early prion phatogenesis. Nat. Med. 2001; 7: 488–492.
  6. Мейл Д. Иммунология. Пер. с англ. Под. ред. А. Ройта, Дж. Бростоффа, Д. Мейла. М.: Мир. 2000. С. 527–546.
  7. D'Orazlo P., Rechnitz G.A. Ion electrode measurements of complement and antibody levels using marker-loaded sheep red blood cell ghosts. Anal. Chem. 1977; 49 (13): 2083–2086.
  8. Haxby J.A., Gotze O., Muller-Eberhard H.J., Kinsky S.C. Release of trapped marker from liposomes by the action of purified complement components. Proc. Natl. Acad. Sci. U.S.A. 1969; 64: 290–298.
  9. Скопинская С.Н., Ярков С.П. Липосомальный иммуноанализ. Иммунология. 1996; 2: 6–12.
  10. Rongen H.A.H., Bult A., van Bennekom W.P. Liposomes and immunoassays. J. Immunol. Meth. 1997; 204: 105–133.
  11. Masaki T., Okada N., Yasuda R., Okada H. Assay of complement activity in human serum using large unilamellar liposomes. J. Immunol. Meth. 1986; 123: 19–24.
  12. Bowden R.W., Rising M., Akot G. et al. Homogeneous, liposome-based assay for total complement activity in serum. Clin. Chem. 1986; 32 (2): 275–278.
  13. Tatsu Y., Yamamura S., Yoshikawa S. Fluorescent fiber-optic immunosensing system based on complement lysis of liposome containing carboxyfluorescein. Biosenssors & Bioelectronics. 1992; 7: 741–745.
  14. Szoka F., Papahadjopoulos D. Procedure for preparation of liposomes with large internal aqueous space and high capture by reverse-phase evaporation. Proc. Nat. Acad. Sci. USA. 1978; 75 (9): 4194–4198.
  15. Скопинская С.Н., Ярков С.П., Храмов Е.Н., Злобин Е.Н. Сульфородамин 101 — новый эффективный маркер в липосомальном иммуноанализе. Вестник РАМН. 2000; 10: 19–24.
  16. Ohsawata T., Miura H., Harada K. Improvement of encapsulation efficiency of water-soluble drugs in liposomes formed by the freeze-thawing method. Chem. Pharm. Bull. 1985; 33 (9): 3945–3952.
  17. Vaskovsky V.E., Kostetsky E.Y., Vasendin I.M. A universal reagent for phospholipids analysis. J. Chromatograph. 1975; 114: 129–141.

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