Immunoassay Using Fluorescent Dye-Trapped Liposomes Liposome Immune Lysis Assay (LILA)
The liposome immune lysis assay (LILA) system was developed by S. C. Kinsky and coworkers in 1968 (Haxby, 1968). They used glucose as an internal liposome marker, and measured the glucose release in an enzymatic reaction. They also introduced umbelliferone phosphate and alkaline phosphatase as the first fluorescent marker system (Six, 1974). The enzyme reaction resulted in both the generation and the amplification of fluorescence. In 1977, Smolarsky et al. developed a novel LILA system using a complex of a fluorescent molecule (1-aminonaphthalene-3,6,8-trisulfonic acid (ANTS)) and a quencher (dipyridinium-p-xylene) as a marker trapped in liposomes. The complex, when entrapped in liposomes, shows little fluorescence. Upon lysis of the liposomes, dilution of the quencher in the external volume leads to a high fluorescence signal. One of the problems of the use of ANTS as a fluorophore is, however, that its excitation and emission wavelengths coincide with those of a fluorescent component existing in complement sources, and precise measurements may not be possible because of an increase in the background fluorescence.
KeywordsAlternative Complement Pathway Sandwich Assay Complement Source Lipid Antigen Glycolipid Antigen
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- Alving, C. R., Shichijo, S. and Mattsby-Baltzer, I. (1984) Preparation and use of liposomes in immunological studies. in “Liposome Technology”, G. Gregoriadis ed., CRC Press, Boca Raton, FL, 2: 157–175.Google Scholar
- Gerlach, E. and Deuticke, B. (1964) Eine einfache Methode zur Mikrobestimmung von Phosphat in der Papierchromatographie. Biochem. Z., 337: 477–479.Google Scholar
- Kabat, E.A. and Mayer, M.M. (1961) “Kabat and Mayer’s Experimental Immunochemistry”, Charles C. Thomas Pub., Springfield, IL, p. 133Google Scholar
- Lelkes, P. I. (1984) Methodological aspects dealing with stability measurements of liposomes in vitro using the carboxyfluorescein. in “Liposome Technology”, G. Gregoriadis ed., CRC Press, Boca Raton, 3: 225–246.Google Scholar
- Ralston, E., Hjelmeland, L. M., Klausner, R. D., Weinstein, J. N. and Blumenthal, R. (1981) Carboxyfluorescein as a probe for liposome-cell interactions: effect of impurities, and purification of the dye. Biochim. Biophys. Acta, 694: 133–137.Google Scholar
- Umeda, M., Ishimori, Y., Yoshikawa, K., Takada, M. and Yasuda, T. (1986b) Liposome immune lysis assay (LILA): Application of sandwich method to determine a serum protein component with antibody bearing liposomes. J. Immunol. Methods, submitted.Google Scholar
- Weinstein, J. N., Ralston, E., Leserman, L. D., Klausner, R. D., Dragsten, P., Henkart, P. and Blumenthal, R. (1984) Self-quenching of carboxyfluorescein fluorescence: uses in studying liposomes stability and liposome-cell interaction. in “Liposome Technology”, G. Gregoriadis ed., CRC Press, Boca Ralton, FL, 3: 183–204.Google Scholar