<p> A novel fabric-based, highly sensitive enzymatic glucose biosensor is reported with dual-sensing technique, integrating both colorimetric and electrochemical methods on a 3D origami platform engineered using eco-sustainable &amp; biodegradable starch-based biopolymer as an additional sensing layer. The study also compares the effectiveness of starch as a much more sensitive coating over traditionally used chitosan with improved LOD. Further, the unique origami design and multi-modal sensing, makes it highly suitable for wearable and POC applications. The 3D origami configuration allows compact folding and fluid manipulation through capillary action, obviating the need for external power sources. The dual-mode detection improves reliability and offers complementary outputs for enhanced diagnostic accuracy demonstrating ultra-high sensitivity with LOD of 10 pM and LOQ of ~ 440 pM for glucose within a linear range of 10 mM to 10 pM for electrochemical and 10 mM to 0.1 µM for colorimetric technique along with a device shelf-life of ~ 30 days under normal conditions. Efficacy of the fabricated glucose sensor has also been tested with real-time human sweat samples under fasting as well as 1&#xa0;h after post breakfast. The major novelty lies in the exploitation of long-chain polymeric groups like starch (polysaccharide) to trap colored iodide complexes formed even at ultra-low glucose concentrations thereby accurately identifying picomolar level of its own monomer viz. glucose (monosaccharide) thereby acting as a potential substitute of chitosan in future devices. Future research will aim to enhance sensor shelf life by improving starch stability against moisture and microbial degradation for prolonged usage. Nevertheless, this innovative approach highlights a promising direction in the development of green, wearable highly sensitive biosensors for continuous and non-invasive health monitoring.</p> Graphical Abstract <p></p>