The gas-phase odor biosensors exhibit outstanding sensitivity and selectivity towards target ligands. Current odor detections, however, were only operated under one or multiple separated olfactory receptors (ORs) stimulated by a single component odor. Meanwhile, the nonlinear characteristic, drift, and aging problems in biosensors were the main barriers for better quantification performance. Here, we proposed a gas-phase odor biosensor for odor mixture quantification based on relative comparison method using cell expressing ORs, the liquid thickness control and liquid exchange maintained repeatable OR responses. We repeatedly compared the known and its adjacent unknown odor response thus mitigating the biosensor drift interference and sensor nonlinearity characteristic. We first conducted the odor quantification of one odor component in a mixture of five odors. The existence of other odors did not influence the quantification of Or56a towards geosmin component in the mixture. Then, two types of OR were utilized for quantifying odor mixture with or without adding the OR label information to the individual cells. With-label quantification was more precise and efficient than without-label one. Lastly, we simultaneously expressed two ORs on one cell line for wider detection range and quantified the mixture even if OR has cross sensitivity. We believe the odor mixture quantification methods established in this study will extend the application scenarios of odor biosensors especially in rapid on-site odor detection.
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