Water-soluble phosphate-functionalized polyfluorene as fluorescence biosensors toward cytochrome c

An anionic water-soluble polyfluorene derivative, poly(9,9-bis(6′-phosphatehexyl)fluorene-alt-1,4-phenylene) sodium salt (PFHPNa), was synthesized by Suzuki coupling reaction in DMF/water. Polymer PFHPNa was well soluble in water with a strong blue fluorescence emission. Effect of the side chain length on fluorescence sensory properties was studied by comparing quenching efficiencies toward different quenchers of PFHPNa with a reported polymer poly(9,9-bis(3′-phosphatepropyl)fluorene-alt-1,4-phenylene) sodium salt (PFPPNa), which have different side chains in length. For small molecular quenchers (methylviologen, MV²⁺) and meso-5,10,15,20-tetrakis-(N-methyl-4-pyridyl)porphine (TMPyP4), polymer PFHPNa had lower sensitivity due to the much longer side chain length. The positively charged metalloprotein cytochrome c could quench fluorescence of conjugated polymers via energy transfer and electron transfer. Moreover, polymer PFHPNa showed higher fluorescence quenching toward large biomolecules than PFPPNa. The corresponding Stern-Volmer (K _sv) value of polymer PFHPNa was determined to be 2.1×10⁸ M⁻¹ for cytochrome c. It could be used as a sensitive and selective fluorescence sensor for protein cytochrome c. Supported by the National Natural Science Foundation of China (Grant Nos. 20574067 & 50633040), the Science Fund for Creative Research Groups (Grant No. 20621401), and the 973 Project (Grant No. 2002CB613402)