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This review describes the development towards actomyosin based nanodevices taking a starting point in pioneering studies in the 1990s based on conventional in vitro motility assays. References are given to parallel developments using the kinesin–microtubule motor system. The early developments focused on achieving cargo-transportation using actin filaments as cargo-loaded shuttles propelled by surface-adsorbed heavy meromyosin along micro- and nanofabricated channels. These efforts prompted extensive studies of surface–motor interactions contributing with new insights of general relevance in surface and colloid chemistry. As a result of these early efforts, a range of complex devices have now emerged, spanning applications in medical diagnostics, biocomputation and formation of complex nanostructures by self-organization. In addition to giving a comprehensive account of the developments towards real-world applications an important goal of the present review is to demonstrate important connections between the applied studies and fundamental biophysical studies of actomyosin and muscle function. Thus the manipulation of the motor proteins towards applications has resulted in new insights into methodological aspects of the in vitro motiliy assay. Other developments have advanced the understanding of the dynamic materials properties of actin filaments.
This review describes the development towards actomyosin based nanodevices taking a starting point in pioneering studies in the 1990s based on conventional in vitro motility assays. References are given to parallel developments using the kinesin–microtubule motor system. The early developments focused on achieving cargo-transportation using actin filaments as cargo-loaded shuttles propelled by surface-adsorbed heavy meromyosin along micro- and nanofabricated channels. These efforts prompted extensive studies of surface–motor interactions contributing with new insights of general relevance in surface and colloid chemistry. As a result of these early efforts, a range of complex devices have now emerged, spanning applications in medical diagnostics, biocomputation and formation of complex nanostructures by self-organization. In addition to giving a comprehensive account of the developments towards real-world applications an important goal of the present review is to demonstrate important connections between the applied studies and fundamental biophysical studies of actomyosin and muscle function. Thus the manipulation of the motor proteins towards applications has resulted in new insights into methodological aspects of the in vitro motiliy assay. Other developments have advanced the understanding of the dynamic materials properties of actin filaments.
Portable biosensor systems would benefit from reduced dependency on external power supplies as well as from further miniaturization and increased detection rate. Systems built around self-propelled biological molecular motors and cytoskeletal filaments hold significant promise in these regards as they are built from nanoscale components that enable nanoseparation independent of fluidic pumping. Previously reported microtubule-kinesin based devices are slow, however, compared to several existing biosensor systems. Here we demonstrate that this speed limitation can be overcome by using the faster actomyosin motor system. Moreover, due to lower flexural rigidity of the actin filaments, smaller features can be achieved compared to microtubule-based systems, enabling further miniaturization. Using a device designed through optimization by Monte Carlo simulations, we demonstrate extensive myosin driven enrichment of actin filaments on a detector area of less than 10 μm², with a concentration half-time of approximately 40 s. We also show accumulation of model analyte (streptavidin at nanomolar concentration in nanoliter effective volume) detecting increased fluorescence intensity within seconds after initiation of motor-driven transportation from capture regions. We discuss further optimizations of the system and incorporation into a complete biosensing workflow.
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