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In this study, a turn-on paper-based optical analytical system with a rapid, sensitive and quantitative response for glucose was developed. The luminescence sensing material, crystalline iridium(III)-Zn(II) coordination polymers, or Ir-Zne, was grown electrochemically on stainless steel mesh and then deposited on filter paper. This sensing substrate was subsequently built up under glucose oxidase encapsulated in hydrogel and then immobilized on egg membrane with the layer-by-layer method. Once the glucose solution was dropped onto the paper, the oxygen content was depleted simultaneously with a concomitant increase in the phosphorescence of Ir-Zne. The detection limit for glucose was 0.05 mM. The linear dynamic range for the determination of glucose was 0.05-8.0 mM with a correlation coefficient ( R 2) of 0.9956 (y=68.11 [glucose]−14.72). The response time was about 0.12 s, and the sample volume was less than 5 μL. The effects of mesh size, buffer concentration, pH, enzyme concentration, temperature, and interference, and the stability of the biosensor, have also been studied in detail. Finally, the biosensor was successfully applied to the determination of glucose in human serum. [ABSTRACT FROM AUTHOR]