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Determination of phycobiliproteins by capillary electrophoresis with laser‐induced fluorescence detection
Author(s) -
Viskari Pertti J.,
Kinkade Christopher S.,
Colyer Christa L.
Publication year - 2001
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(20017)22:11<2327::aid-elps2327>3.0.co;2-j
Subject(s) - phycobiliprotein , allophycocyanin , phycocyanin , capillary electrophoresis , chemistry , phycobilisome , phycoerythrin , fluorescence , sodium dodecyl sulfate , chromatography , absorbance , laser induced fluorescence , chromophore , cyanobacteria , analytical chemistry (journal) , photochemistry , biology , genetics , flow cytometry , physics , quantum mechanics , bacteria
Phycobiliproteins are derived from the photosynthetic apparatus of cyanobacteria and eukaryotic algae. They are composed of a protein backbone to which linear tetrapyrrole chromophores are covalently bound. Furthermore, they are water‐soluble, highly fluorescent, and relatively stable at room temperature and neutral pH. For this reason, capillary electrophoresis‐laser induced fluorescence (CE‐LIF) seems the ideal method for determination of these important proteins. The effects of buffer additives such as sodium dodecyl sulfate (SDS)and putrescine on the separation of the three major phycobiliprotein types, namely allophycocyanin, phycocyanin, and phycoerythrin, with excitation and emission maxima at 652/660, 615/647, and 565(494)/575 nm, respectively, are considered. Detection limits for these proteins by CE‐LIF are some 60–500 times better than by absorbance detection. The development of a fast and sensitive CE‐LIF assay such as this is of potential significance to our understanding of chemical and biological oceanographic processes.