Many carbon nanotube field-effect transistor (CNT-FET) studies have used immobilized antibodies as the ligand binding moiety. However, antibodies are not optimal for CNT-FET detection due to their large size and charge. Their size can prevent ligands from reaching within the Debye length of the CNTs and a layer of charged antibodies on the circuits can drown out any ligand signal. In an attempt to minimize the antibody footprint on CNT-FETs, we examined whether pyrene hydrazide modification of antibody carbohydrates could reduce the concentration required to functionalize CNT circuits. The carbohydrates are almost exclusively on the antibody Fc region and this site-specific modification could mediate uniform antibody orientation on the CNTs. We compared the hydrazide modification of anti-E. coliO157:H7 polyclonal antibodies to pyrenebutanoic acid succinimidyl ester-coated CNTs and carbodiimide-mediated antibody CNT attachment. Our results show that the pyrene hydrazide modification was superior to those methods with respect to bacteria detection and less than 1 nM labeled antibody was required to functionalize the circuits.

Biomolecular detections performed on carbon nanotube field-effect transistors (CNT-FETs) frequently use reactive pyrenes as an anchor to tether bioactive ligands to the hydrophobic nanotubes. In this paper, we explore the possibility of directly using bioactive aromatic compounds themselves as CNT-FET ligands. This would be an efficient way to functionalize CNT-FETs since many aromatic compounds bind avidly to nanotubes, and it would also ensure that ligand-binding molecules would be brought in close proximity to the nanotubes. Using a model system consisting of pyrene, phenanthrene, naphthalene, or phenyl boronic acids immobilized on CNT-FET wafers, we show that all are able to bind glycated human serum albumin (gHSA), which is an important diabetes marker. Pyrene boronic acid proved to bind CNTs with the greatest apparent affinity as measured by gHSA impedance. Interestingly, gHSA CNT-FET signal intensity, which is proportional to amount of protein bound, remained essentially unchanged for all the boronic acids tested.

Label-free and real-time detection technologies can dramatically reduce the time and cost of pharmaceutical testing and development. However, to reach their full promise, these technologies need to be adaptable to high-throughput automation. To demonstrate the potential of single-walled carbon nanotube field-effect transistors (SWCNT-FETs) for high-throughput peptide-based assays, we have designed circuits arranged in an 8 × 12 (96-well) format that are accessible to standard multichannel pipettors. We performed epitope mapping of two HIV-1 gp160 antibodies using an overlapping gp160 15-mer peptide library coated onto nonfunctionalized SWCNTs. The 15-mer peptides did not require a linker to adhere to the non-functionalized SWCNTs, and binding data was obtained in real time for all 96 circuits. Despite some sequence differences in the HIV strains used to generate these antibodies and the overlapping peptide library, respectively, our results using these antibodies are in good agreement with known data, indicating that peptides immobilized onto SWCNT are accessible and that linear epitope mapping can be performed in minutes using SWCNT-FET.