Journal
CARBOHYDRATE RESEARCH
Volume 346, Issue 14, Pages 2156-2164Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.carres.2011.07.014
Keywords
Polysaccharide; Fluorescent; Labeling; Fluorescein-5-thiosemicarbazide; Imaging; Living cells
Funding
- National Natural Science Foundation of China (NSFC) [30870548, 31071506]
- Foundation of Key Laboratory of Resource Biology and Biotechnology in Western China
- Ministry of Education in Northwest University [KH09019]
- Hi-Tech Research and Development Program of China [2007AA10Z338]
- Program for New Century Excellent Talents in Universities [NCET-08-0893]
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A simple and efficient procedure for the fluorescent labeling of saccharides is a prerequisite step for imaging the transport of polysaccharides in living cells. We report a one-pot strategy for the fluorescent labeling of saccharides with fluorescein-5-thiosemicarbazide (FTSC), which introduces the thiosemicarbazide group of FTSC to the aldehyde group at the reducing end of saccharides to form stable amino derivatives via reductive amination. The Glc-FTSC derivative was characterized by HPLC-MS, HRESIMS and NMR spectroscopy. Saccharides were quantitatively labeled with FTSC at 75 degrees C for 1 h under optimum reaction conditions. Fluorescence studies illustrated that the conjugation of FTSC to saccharides did not change its florescence properties (lambda(ex) = 495 nm, lambda(em) = 517 nm), presenting desirable compatibility with commonly used fluorescence equipment. Polysaccharide AAG-FTSC derivatives exhibited rather low levels of cytotoxicity against rat thymus cells, and the fluorescent labeling procedure had slight impact on their anti-tumor activity. Results indicate that the assay neither introduces discernible cytotoxicity against living cells nor obviously alters the functional activities of polysaccharides, and provides a convenient, highly efficient fluorescent labeling approach for imaging the transport of polysaccharides in living cells. (C) 2011 Elsevier Ltd. All rights reserved.
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