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Biomedical application of metalloporphyrins room-temperature phosphorescence
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Paper Abstract

Biological specimens are characterized by high light-scattering and background fluorescence which cause decrease in sensitivity of immunoassays. These unfavorable effects can be minimized by selecting a label with high absolute sensitivity, appropriate excitation and emission wavelengths, and time-resolved mode of measurements. This can be achieved by the use of metalloporphyrin labels, microsecond logic for discrimination of scattered light and background fluorescence which arises in optical elements, cells, solvents, and samples. The intensity of metalloporphyrin phosphorescence depends to a great extent on pH. Nonionic and cationic micelles have a strong effect on the phosphorescence quantum yield of Pd- coproporphyrin. Under optimum conditions Pd-coproporphyrin can be detected at a concentration of 10-13M by using modified Arcus 1230 (Wallac, Finland). On the basis of these results a novel class of luminescent labels --phosphorescent metalloporphyrins -- was applied to immunoassays. Pd- and Pt-coproporphyrins were used for the covalent labeling of antibodies and antigens. Special derivatives of a porphyrin with activated side chain were synthesized. Techniques for covalent coupling of porphyrins and their metal derivatives with proteins were developed as well as methods for purification of conjugates. A solid-phase time-resolved porphyrin phosphorescence immunoassay test was developed. The well known `sandwich' and competition techniques are compatible with the proposed method. Two or more metalloporphyrins with distinguishable phosphorescent parameters may be used for the simultaneous determining of several antigens in one sample.

Paper Details

Date Published: 18 May 1993
PDF: 11 pages
Proc. SPIE 1885, Advances in Fluorescence Sensing Technology, (18 May 1993); doi: 10.1117/12.144705
Show Author Affiliations
Alexander P. Savitsky, Bach Institute of Biochemistry (Russia)

Published in SPIE Proceedings Vol. 1885:
Advances in Fluorescence Sensing Technology
Joseph R. Lakowicz; Richard B. Thompson, Editor(s)

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