Nanofluidic concentration of selectively extracted biomolecule analytes by microtubules

We present a novel device for the selective extraction and high concentration of biomolecule analytes by integrating microtubules, one of cytoskeletal filaments, with nanofluidic technologies. Microtubules can be functionalized to provide numerous, nanoscopic binding sites for specific target biomolecules. The functionalized microtubules, with the target biomolecules bound to their surface, can be transported in the opposite direction to nontarget molecules using electrokinetic separation. Subsequently, the target molecule-bound microtubules are concentrated by a flat nanochannel structure, which filters the microtubules but allows ionic current and flow to pass through it. This device makes it possible to selectively extract target molecules such as streptavidin and bovine serum albumin and then highly concentrate them up to higher than 5 orders of magnitude from a complex mixture of analytes ranging from 1 nM to 10 fM. In addition, the device performs both extraction (separation) and concentration process simultaneously, which are typically performed in order in other devices, so that we significantly reduce analysis time and labor and even enable preconcentrated, identified target molecules to be available for postanalysis. Thus, we believe that the use of functionalized microtubules with nanofluidics will be a useful means to facilitate biochemical analysis systems.