A novel amperometric glucose biosensor based on self-assembling glucose oxidase (GOx) and dendrimer-encapsulated Pt nanoparticles (Pt-DENs) on nanofibrous polyaniline (PANI) was described. PANI nanofibers were synthesized via an interfacial polymerization method. A sulfonated polyelectrolytes-poly(sodium 4-styrenesulfonate) (PSS) was used to form the negative PANI/sulfonated polyelectrolyte complex, which had good disperse in aqueous solution. GOx was immobilized on the PANI/PSS surface by alternatively assembling a cationic Pt-DENs layer and an anionic GOx layer. The unique sandwich-like layer structure (Pt-DENs/GOx/Pt-DENs/PANI/PSS) formed by self-assembling provides a favorable microenvironment to keep the bioactivity of GOx and to prevent enzyme molecule leakage. The fabricated Pt-DENs/GOx/Pt-DENs/PANI/ PSS electrode exhibited excellent response performance to glucose with a detection limit of 0.5 mu M, wide linear range from 10 mu M to 4.5 mM, short response time within 5 s, improved sensitivity of 39.63 mu A/(mM cm(2)), and good stability (85% remains after 20 days). (C) 2008 Wiley Periodicals, Inc.