A Sensitive Capacitive Biosensor for Protein a Detection Using Human IgG Immobilized on an Electrode Using Layer-by-Layer Applied Gold Nanoparticles

A capacitive biosensor for the detection of protein A was developed. Gold electrodes were fabricated by thermal evaporation and patterned by photoresist photolithography. A layer-by-layer (LbL) assembly of thiourea (TU) and HAuCl4 and chemical reduction was utilized to prepare a probe with a different number of layers of TU and gold nanoparticles (AuNPs). The LbL-modified electrodes were used for the immobilization of human IgG. The binding interaction between human IgG and protein A was detected as a decrease in capacitance signal, and that change was used to investigate the correlation between the height of the LbL probe and the sensitivity of the capacitive measurement. The results showed that the initial increase in length of the LbL probe can enhance the amount of immobilized human IgG, leading to a more sensitive assay. However, with thicker LbL layers, a reduction of the sensitivity of the measurement was registered. The performance of the developed system under optimum set-up showed a linearity in response from 1 × 10−16 to 1 × 10−13 M, with the limit detection of 9.1 × 10−17 M, which could be interesting for the detection of trace amounts of protein A from affinity isolation of therapeutic monoclonal antibodies.

Growth and Drug Interaction Monitoring of NIH 3T3 Cells by Image Analysis and Capacitive Biosensor

Capacitive biosensors are manufactured on glass slides using the semiconductor process to monitor cell growth and cell–drug interactions in real time. Capacitance signals are continuously monitored for each 10 min interval during a 48 h period, with the variations of frequency from 1 kHz to 1 MHz. The capacitance values showed a gradual increase with the increase in NIH 3T3 cell numbers. After 48 h of growth, 6.67 μg/mL puromycin is injected for the monitoring of the cell–drug interaction. The capacitance values rapidly increased during a period of about 10 h, reflecting the rapid increase in the cell numbers. In this study, we monitored the state of cells and the cell–drug interactions using the developed capacitive biosensor. Additionally, we monitored the state of cell behavior using a JuLiTM Br&FL microscope. The monitoring of cell state by means of a capacitive biosensor is more sensitive than confluence measuring using a JuLiTM Br&FL microscope image. The developed capacitive biosensor could be applied in a wide range of bio-medical areas; for example, non-destructive real-time cell growth and cell–drug interaction monitoring.

Microfluidic integrated capacitive biosensor for C-Reactive Protein label-free and real-time detection

Microfluidic chip has been integrated with capacitive biosensor based on mass-producible three-dimensional (3D) interdigital electrode arrays. To achieve the monitoring of biosensor preparation and the cardiac and periodontitis-related biomarkers, all...

A Probeless Capacitive Biosensor for Direct Detection of Amyloid Beta 1-42 in Human Serum Based on an Interdigitated Chain-Shaped Electrode

Amyloid beta (aβ) 1-42, a peptide that is 1-42 amino acids long, is a major component of senile plaques in the brains of patients with Alzheimer’s disease. Aβ detection has become an essential antecedence to predict the declining mental abilities of patients. In this paper, a probeless capacitive biosensor for the non-Faradaic detection of aβ 1-42 peptide was developed by immobilizing a specific anti-aβ antibody onto a self-assembled monolayer functionalized interdigitated chain-shaped electrode (anti-aβ/SAM/ICE). The novelty and difference of this article from previous studies is the direct detection of aβ peptide with no redox probe ((Fe(CN)6)3−/4−), which can avoid the denaturation of the protein caused by the metallization (binding of aβ to metal ion Fe which is presented in the redox couple). The direct detection of aβ with no redox probe is performed by non-Faradaic capacitive measurement, which is greatly different from the Faradaic measurement of the charge transfer resistance of the redox probe. The detection of various aβ 1-42 peptide concentrations in human serum (HS) was performed by measuring the relative change in electrode interfacial capacitance due to the specific antibody-aβ binding. Capacitance change in the anti-aβ/SAM/ICE biosensor showed a linear detection range between 10 pg mL−1 and 104 pg mL−1, and a detection limit of 7.5 pg mL−1 in HS, which was much lower than the limit of detection for CSF aβ 1-42 (~500 pg mL−1) and other biosensors. The small dissociation constant Kd of the antibody-antigen interaction was also found to be 0.016 nM in HS, indicating the high binding affinity of the anti-aβ/SAM/ICE biosensor in the recognizing of aβ 1-42. Thus, the developed sensor can be used for label-free and direct measurement of aβ 1-42 peptide and for point-of-care diagnosis of Alzheimer’s disease without redox probe.