Acid-Modulated Peptide Synthesis for Application on Oxide Biosensor Interfaces.

In this paper we report an acid-modulated strategy for novel peptide microarray production on biosensor interfaces. We initially selected a controlled pore glass (CPG) as a support for solid-phase peptide synthesis (SPPS) to implement a chemistry that can be performed at the interface of multiple field effect transistor (FET) sensors, eventually to generate label-free peptide microarrays for protein screening. Our chemistry uses a temporary protection of the N-terminal amino function of each amino acid building block with a tert-butyloxycarbonyl (Boc) group that can be removed after each SPPS cycle, in combination with semi-permanent protection of the side chains of trifunctional amino acid residues. Such a protection scheme with a well-proven record of application in conventional, batchwise SPPS has been fine-tuned for optimal performance on CPG and, from there, translated to SPR chips that allow layer-by-layer monitoring of amino acid coupling. Our results validate this acid-modulated synthesis as a feasible approach for producing peptides in high yields and purity on flat glass surfaces, such as those in bio-FETs.

Planar Junctionless Field-Effect Transistor for Detecting Biomolecular Interactions.

Label-free field-effect transistor-based immunosensors are promising candidates for proteomics and peptidomics-based diagnostics and therapeutics due to their high multiplexing capability, fast response time, and ability to increase the sensor sensitivity due to the short length of peptides. In this work, planar junctionless field-effect transistor sensors (FETs) were fabricated and characterized for pH sensing. The device with SiO2 gate oxide has shown voltage sensitivity of 41.8 ± 1.4, 39.9 ± 1.4, 39.0 ± 1.1, and 37.6 ± 1.0 mV/pH for constant drain currents of 5, 10, 20, and 50 nA, respectively, with a drain to source voltage of 0.05 V. The drift analysis shows a stability over time of -18 nA/h (pH 7.75), -3.5 nA/h (pH 6.84), -0.5 nA/h (pH 4.91), 0.5 nA/h (pH 3.43), corresponding to a pH drift of -0.45, -0.09, -0.01, and 0.01 per h. Theoretical modeling and simulation resulted in a mean value of the surface states of 3.8 × 1015/cm2 with a standard deviation of 3.6 × 1015/cm2. We have experimentally verified the number of surface sites due to APTES, peptide, and protein immobilization, which is in line with the theoretical calculations for FETs to be used for detecting peptide-protein interactions for future applications.

Nanoimmunosensor based on ZnO nanorods for ultrasensitive detection of 17ß-Estradiol.

Advances in nanostructured materials have facilitated the development of novel sensitive techniques for detection of environmental and clinical analytes. There is immense need for development of devices that can detect analytes at concentrations as low as few pg mL-1. The comparable size of nanostructured materials and biomolecules enabled the integration of biological systems with nanometer sized structures. Herein, we demonstrate a Zinc Oxide nanorods (ZnONRs) integrated ultrasensitive label-free biosensor with femtomolar (0.01 pg mL-1) sensitivity for the endocrine disruptor 17ß-Estradiol (E2). The ZnONRs, average width 50 nm and length 325 nm, were grown on the silver electrode surface (Ag-ZnONRs). Monoclonal antibodies of E2 (mAb-E2) were covalently immobilized on ZnONRs surface and measured using electrochemical impedance spectroscopy (EIS). A linear detection range of 0.1-200 pg mL-1 for E2 with R2 = 0.99 and % RSD = 4.35 (n = 3, assay volume 90 µL) was achieved for the developed nano-sensing system. A significant enhancement in the sensitivity was achieved in the presence of ZnONRs, enabling the limit of quantification down to 0.1 pg mL-1 with 2.7 % capacitance change per decade. In addition, a further increase in sensitivity due to assay volume reduction (20 µL) was observed enabling further scope of miniaturization.