title: One-pot fluorescent labeling of saccharides with fluorescein-5-thiosemicarbazide for imaging polysaccharides transported in living cells.
abstract: 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°C for 1 h under optimum reaction conditions. Fluorescence studies illustrated that the conjugation of FTSC to saccharides did not change its florescence properties (λ(ex)=495 nm, λ(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.
authors.affiliation: Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Science, Northwest University, 229 Northern Taibai Road, Xi'an 710069, PR China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.forename: Zhongfu
authors.initials: Z
authors.lastname: Wang
authors.forename: Xiaorui
authors.initials: X
authors.lastname: Zhang
authors.forename: Wenxia
authors.initials: W
authors.lastname: Zhou
authors.forename: Linjuan
authors.initials: L
authors.lastname: Huang
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism / Q000494:pharmacology / Q000633:toxicity; D001692:Biological Transport; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D005452:Fluoresceins / Q000737:chemistry / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D051379:Mice; D057054:Molecular Imaging / Q000379:methods*; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology / Q000633:toxicity; D051381:Rats; D013950:Thymus Gland / Q000166:cytology / Q000187:drug effects
chemical_list: D000970:Antineoplastic Agents; D005452:Fluoresceins; D005456:Fluorescent Dyes; D011134:Polysaccharides; C027912:fluorescein thiosemicarbazide; C008315:maltodextrin