Study and optimization of the selectivity of an odorant binding protein-based bioelectronic nose

Over the past two decades, the use of odorant-binding proteins (OBPs) for the development of biosensors and bioelectronic noses (bioeNs) aimed at detecting and analyzing volatile organic compounds (VOCs) has been the subject of considerable research. However, there is a lack of fundamental studies for better understanding the interaction between OBPs and VOCs in gas phase. In this work, we investigated the effect of two key factors, namely relative humidity (RH) level and immobilization technique, on the selectivity of two OBP-based biosensors in gas phase. Concerning the effect of RH, the results showed that our active OBP (wild-type rat OBP3) lost its selectivity at 0% RH but retained good selectivity at 30% and 50% RH. To better understand the effect of this parameter, the hydration mechanism of the OBP was studied both experimentally and through molecular dynamics simulations. The effect of a cysteine residue, genetically added to the N-terminus of OBPs to control their orientation after immobilization on the chip, was evaluated. A significant reduction in selectivity was observed in the absence of cysteine. As expected, the introduction of this amino acid enabled to control the orientation of OBPs, making their binding pocket more accessible to VOCs and favoring specific interactions. Furthermore, we demonstrated that combining OBP-based biosensors with different properties can improve the discrimination capability of our bioeN. Finally, the ability of our system to detect essential oil vapors was tested, providing preliminary evidence that our bioeN is capable of detecting VOCs in complex media.