Photophysical Features of Protolytic Equilibria of Fluorescein Bifluorophores

IF 0.8 4区化学 Q4 SPECTROSCOPY

Journal of Applied Spectroscopy Pub Date : 2024-11-19 DOI:10.1007/s10812-024-01820-x

F. Fan, V. A. Povedailo, I. L. Lysenko, O. L. Sharko, I. O. Mazunin, S. A. Tikhomirov, V. V. Shmanai

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Abstract

The effects of pH and the solution composition on the photophysical properties of two carboxyfluorescein bifluorophores were studied along with a comparison of the fluorescence effect of these bifluorophores with that of monomer carboxyfluoresceins (5-FAM and 6-FAM) in conjugates with protein molecules. Both the bifluorophores were found to fluoresce well in an alkaline medium but the fluorescence intensity decreased at pH 6–7 and almost disappeared in acidic media. The fluorescence of meta-(6-FAM)₂ is very sensitive to the solution composition, while the fluorescence of meta-(5-FAM)₂ is more stable and depends only slightly on the solution properties. Upon formation of a conjugate of the bifluorophore with a protein, the fluorescence yield of the bifluorophore decreases to a greater extent in comparison with the FAM monomers but the total fluorescence yield is not less than the intensities of the two carboxyfluorescein monomer–protein conjugates due to the high extinction coefficient of the bifluorophore (~140,000 M^–1·cm^–1).

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荧光素双荧光团原生溶解平衡的光物理特征

研究了 pH 值和溶液成分对两种羧基荧光素双荧光团光物理性质的影响，并比较了这些双荧光团与单体羧基荧光素（5-FAM 和 6-FAM）在与蛋白质分子共轭时的荧光效果。结果发现，这两种双荧光团在碱性介质中都能发出良好的荧光，但在 pH 值为 6-7 时荧光强度下降，在酸性介质中几乎消失。元-(6-FAM)2 的荧光对溶液成分非常敏感，而元-(5-FAM)2 的荧光则更为稳定，对溶液性质的影响很小。当双荧光团与蛋白质形成共轭物时，与 FAM 单体相比，双荧光团的荧光产率会有较大程度的下降，但由于双荧光团的消光系数较高（约 140,000 M-1-cm-1），其总荧光产率不会低于两种羧基荧光素单体-蛋白质共轭物的强度。

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来源期刊

Journal of Applied Spectroscopy SPECTROSCOPY-

CiteScore

1.30

自引率

14.30%

发文量

145

审稿时长

2.5 months

期刊介绍： Journal of Applied Spectroscopy reports on many key applications of spectroscopy in chemistry, physics, metallurgy, and biology. An increasing number of papers focus on the theory of lasers, as well as the tremendous potential for the practical applications of lasers in numerous fields and industries.