cAMP-specific 3′,5′-cyclic phosphodiesterase 4B (PDE4B) was found to constitute an excellent therapeutic target. Inhibition of PDE4B suppresses inflammation33. Roliparm and roflumilast were reported as potential PDE4B inhibitors for the treatment of inflammation34–36. Although different methods, including, ELISA, gene cloning and radioisotope/alumina acid methods, are available to screen PDE4B inhibitors, these methods are still highly time-consuming. For example, cloning usually takes more than two weeks to establish PDE4B gene-expressing vectors. Consequently, even more time is required for compound screening. Likewise, to perform drug screening using radioisotope/alumina acid methods, handling a liquid system and preparation of alumina plates is inconvenient37–40. In comparison, the inkjet printing-based screening approach offers a user-friendly method and is much less time-consuming. The fundamental strategy in this work utilizes non-fabricated parchment paper as a substrate to sense PDE4B inhibitor activity, and in turn, this sensing event produces FRET signals. We present an alternative system, which will substantially enhance the ability to screen future FRET signal-based assay for novel analyses. Our study involves paper-based inkjet bioprinting for FRET signal determination via evaluating the competition between Eu-labelled cAMP tracer and cAMP for binding sites on cAMP- specific mAb labeled with ULight. After printing the reaction components on parchment paper, antibodies bound to the Eu-labelled cAMP. When Eu-chelate molecules were excited at 320/340 nm, the energy is transferred by FRET to ULight molecules, which in turn emitted fluorescence at 665 nm. cAMP-specific PDE4B inhibition results in the production of FRET-based antibody interaction, which displayed a uniquely characteristic low-background signal on parchment paper. Therefore, FRET signals determined at a lower optical background enables us to analyze threshold values sensitively. The experimental results clearly confirm the compatibility and applicability of this inkjet-printing system for quantitative FRET determination. Previous studies have reported an inkjet printing-based multi-enzyme printing system (ADH–DP), to print enzyme and substrate on a piece of parchment paper for certain applications, such as biosensing9. Our previous study proposed a novel concept IM50 for the quantitative determination of inhibitor efficacy using a conventional inkjet printer containing four cartridges32. In addition, earlier reports have verified that gradient color codes preset on a computer can be utilized to quantitatively print enzymes, proteins, and inhibitors on paper. This basic background allowed us to predict the number of mole of component used to react with enzymes/inhibitors. It was also found that a controlled spot size and pattern design played critical roles in creating a paper-based enzymatic assay9,32.