Real time protein recognition in a liquid-gated carbon nanotube field-effect transistor modified with aptamers.

The combination of optimized and passivated Field Effect Transistors (FETs) based on carbon nanotubes (CNTs) together with the appropriate choice and immobilization strategy of aptamer receptors and buffer concentration have allowed the highly sensitive and real time biorecognition of proteins in a liquid-gated configuration. Specifically we have followed the biorecognition process of thrombin by its specific aptamer. The aptamer modified device is sensitive enough to capture a change in the electronic detection mechanism, one operating at low protein concentrations and the other in a higher target concentration range. The high sensitivity of the device is also sustained by the very low detection limits achieved (20 pM) and their high selectivity when other target proteins are used. Moreover, the experimental results have allowed us to quantify the equilibrium constant of the protein-aptamer binding and confirm its high affinity by using the Langmuir equation.

Interaction of biomolecules sequentially deposited at the same location using a microcantilever-based spotter.

A microspotting tool, consisting of an array of micromachined silicon cantilevers with integrated microfluidic channels is introduced. This spotter, called Bioplume, is able to address on active surfaces and in a time-contact controlled manner picoliter of liquid solutions, leading to arrays of 5 to 20-microm diameter spots. In this paper, this device is used for the successive addressing of liquid solutions at the same location. Prior to exploit this principle in a biological context, it is demonstrated that: (1) a simple wash in water of the microcantilevers is enough to reduce by >96% the cross-contamination between the successive spotted solutions, and (2) the spatial resolution of the Bioplume spotter is high enough to deposit biomolecules at the same location. The methodology is validated through the immobilization of a 35mer oligonucleotide probe on an activated glass slide, showing specific hybridization only with the complementary strand spotted on top of the probe using the same microcantilevers. Similarly, this methodology is also used for the interaction of a protein with its antibody. Finally, a specifically developed external microfluidics cartridge is utilized to allow parallel deposition of three different biomolecules in a single run.