Multiplex transport and detection of cytokines using kinesin-driven molecular shuttles

The application of biomolecular active transport systems offers a potential route for downscaling multiple analyte assays for lab-on-a-chip applications. Recently, the capture and transport of a wide range of target analytes including proteins, virus particles, and DNA have been demonstrated using kinesin-driven molecular shuttles. The molecular shuttles consisted of microtubule (MT) filaments that were functionalized with either analyte-selective antibodies or complementary DNA, thus facilitating selective target capture and transport. In the present work, we have applied this microfluidic platform for the simultaneous detection of multiple target protein analytes. Multiplexing of molecular shuttles was achieved by immobilizing biotinylated antibodies against interleukin-2 (IL-2) and tumor necrosis factor-alpha (TNF-alpha) on biotinylated MTs using a streptavidin bridge. Nanocrystal quantum dots of different sizes and spectral emissions were functionalized with IL-2 and TNF-alpha antibodies to facilitate multiplexed detection. In this paper we discuss the results of selectivity and motility in single and multiplexed assays.