Synthesis and characterization of fluorescent perylene bisimide-containing glycopolymers for Escherichia coli conjugation and cell imaging

Abstract

Fluorescent glycopolymers were prepared via combined atom transfer radical polymerization (ATRP) and ‘Click Chemistry’ in one-pot synthesis, in the presence of 2-azidoethyl methacrylate (AzEMA), 2-propynyl α- d-mannopyranoside, N, N′-bis{2-[2-[(2-bromo-2-methylpropanoyl)oxy]ethoxy]ethyl}perylene-3,4,9,10-tetracarboxylic acid bisimide (PBI-Br), copper (I) bromide catalyst and pentamethyldiethylenetriamine (PMDETA) ligand. Simultaneous ATRP and ‘Click Chemistry’ is an attractive method for the synthesis of functional glycopolymers as the reaction conditions are compatible with ATRP of an azide monomer, as long as an alkynyl-functionalized carbohydrate is available for click coupling reaction. The fluorescent glycopolymers were characterized by 1H NMR, FT-IR, UV–visible absorption, fluorescence and X-ray photoelectron spectroscopies, as well as by gel permeation chromatography. Incubation of Escherichia coli (E. coli DH5 α) with the fluorescent glycopolymers yielded green fluorescent bacterial clusters. The low cytotoxicity level of the fluorescent glycopolymers was revealed by incubation with 3T3 fibroblasts, macrophages and KB cells in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays in vitro. Subsequently, the water-soluble, biocompatible and fluorescent glycopolymers were used as effective fluorescent cell labeling agents.