In this project, we have developed a cost-effective nanofluidic-based immunosensors for kinetic measurement of fluorescent-labelled protein binding. With the combination of fluorescence microscopy and reversed buffer flow operation, association and dissociation kinetics can be accessed in one single experiment without extra buffer loading step. Kinetic constants of two representative protein-ligand binding pairs (streptavidin-biotin; IgG/anti-IgG) were quantified. The good agreement of extracted rate constants with literature values and analogous SPR measurements indicates that this approach is applicable to study protein interactions of medium- and high-affinities with a limit of detection down to 1 pM, regardless of the analyte size.
- M. Pugnière, Institut de Recherche en Cancérologie de Montpellier, INSERM
- C.F. Chou, Institute of Physics, Academia Sinica (Taiwan)
Selective list of publication:
- T. Leïchlé, Y.-L. Lin, P.-C. Chiang, S.-M. Hu, K.-T. Liao, and C.-F. Chou, "Biosensor-compatible encapsulation for pre-functionalized nanofluidic channels using asymmetric plasma treatment", Sensors and Actuators B, 161 (1), pp. 805-810 (2012)
- T. Leïchlé, and C.-F. Chou, "Biofunctionalized nanoslits for wash-free and spatially resolved real-time sensing with full target capture", Biomicrofluidics, 9, 034103 (2015)
- P. Teerapanich, M. Pugnière, C. Henriquet, Y.-L. Lin, C.-F. Chou, and T Leïchlé, "Nanofluidic Fluorescence Microscopy (NFM) for real-time monitoring of protein binding kinetics and affinity studies", Biosensors & Bioelectronics, doi.org/10.1016/j.bios.2016.06.033 (2016)