Dawoon Han

Analytical detection of biological thiols in a microchip capillary channel.

Sulfur-containing amino acids, such as cysteine and homocysteine play crucial roles in biological systems for the diagnosis of medical states. In this regard, this paper deals with separation, aliquot and detection of amino thiols on a microchip capillary electrophoresis with electrochemical detection in an inverted double Y-shaped microchannel. Unlike the conventional capillary electrophoresis, the modified microchannel design helps in storing the separated thiols in different reservoirs for further analysis, if required; and also eliminates the need of electrodes regeneration. The device was fabricated using conventional photolithographic technique which consisted of gold microelectrodes on a soda lime glass wafer and microchannels in PDMS mold. Multiple detections were performed using in-house fabricated dual potentiostat. Based on amperometric detection, cysteine and homocysteine were analyzed in 105 s and 120 s, respectively after diverting in branched channels. Repeated experiments proved the good reproducibility of the device. The device produced a linear response for both cysteine and homocysteine in electrochemical analysis. To prove the practicality of device, we also analyzed cysteine and homocysteine in real blood samples without any pre-treatment. Upon calculation, the device showed a very low limit of detection of 0.05 μM. The modified microchip design shall find a broad range of analytical applications involving assays of thiols and other biological compounds.

Microscale loop-mediated isothermal amplification of viral DNA with real-time monitoring on solution-gated graphene FET microchip.

Rapid and reliable molecular analysis of DNA for disease diagnosis is highly sought-after. FET-based sensors fulfill the demands of future point-of-care devices due to its sensitive charge sensing and possibility of integration with electronic instruments. However, most of the FETs are unstable in aqueous conditions, less sensitive and requires conventional Ag/AgCl electrode for gating. In this work, we propose a solution-gated graphene FET (SG-FET) for real-time monitoring of microscale loop-mediated isothermal amplification of DNA. The SG-FET was fabricated effortlessly with graphene as an active layer, on-chip co-planar electrodes, and polydimethylsiloxane-based microfluidic reservoir. A linear response of about 0.23V/pH was seen when the buffers from pH 5-9 were analyzed on the SG-FET. To evaluate the performance of SG-FET, we monitored the amplification of Lambda phage gene as a proof-of-concept. During amplification, protons are released, which gradually alters the Dirac point voltage (VDirac) of SG-FET. The resulting device was highly sensitive with a femto-level limit of detection. The SG-FET could easily produce a positive signal within 16.5min of amplification. An amplification of 10ng/μl DNA for 1h produced a ∆VDirac of 0.27V. The sensor was tested within a range of 2×102 copies/μl (10 fg/μl) to 2×108 copies/μl (10ng/μl) of target DNA. Development of this sensing technology could significantly lower the time, cost, and complications of DNA detection.

Highly selective organic transistor biosensor with inkjet printed graphene oxide support system

Most of the reported field effect transistors (FETs) fall short of a general method to uniquely specify and detect a target analyte. For this reason, we propose a pentacene-based FET with a graphene oxide support system (GOSS), composed of functionalized graphene oxide (GO) ink. The GOSS with a specific moiety group to capture the biomaterial of interest was inkjet printed on the pentacene FET. It provided modular receptor sites on the surface of pentacene, without alteration of the device. To evaluate the performance of a GOSS-pentacene FET biosensor, we detected the artificial DNA and circulating tumor cells as a proof-of-concept. The mobility of the FET dramatically changed upon capturing the target biomolecule on the GOSS. The FET exhibited high selectivity with 0.1 pmoles of the target DNA and a few cancer cells per detection volume. This study suggests a valuable sensor for medical diagnosis that can be mass produced effortlessly at low-cost.

Rapid Detection of Protein Kinase on Capacitive Sensing Platforms

In this study, we developed a capacitive sensor for the one-step and label-free detection of protein kinase A (PKA) enzyme. Metal-insulator-semiconductor (MIS) and electrolyte-insulator-semiconductor (EIS) are a simple electronic transducer, which allows efficient detection of the target analyte. For this reason, we performed a comparative sensing of PKA on the MIS and EIS capacitive sensor. The PKA-specific aptamer was used for the one-step detection. For the immobilization of thiolated aptamer, the MIS sensor contained a thin gold layer, whereas the EIS sensor had a self-aligned monolayer of gold nanoparticles. The interaction of aptamer and PKA changed the charge and density of the sensor surface. The quantitative detection of PKA was performed by analyzing the capacitance-voltage curve after the aptamer-PKA interaction. The MIS and EIS sensor showed a detection limit of 5 U/mL and 1 U/mL, respectively, for the detection of PKA. This study suggests valuable sensing platforms for the rapid and sensitive biochemical diagnosis.

Screening and electrochemical detection of an antibiotic producing gene in bacteria on an integrated microchip

We demonstrate the electrochemical determination of a secondary metabolite producing gene on an integrated microfluidic chip. The fabricated microchip was assembled with a continuous channel for polymerase chain reaction (PCR) and an electrochemical detector in order to achieve rapid and sensitive determination of the valC gene. valC is a gene responsible for producing antibiotic validamycin A in Streptomyces hygroscopicus. Biotin-conjugated primers amplified the valC gene. After the PCR, a DNA amplicon was analyzed in the electrochemical cell containing a streptavidin functionalized Au working electrode. The guanosine present in the DNA amplicon released electrons upon electrochemical oxidation at 0.93 V and the peak current linearly increased with the concentration of the captured DNA amplicon. The fabricated chip successfully amplified and detected the valC gene as low as 30 pg μL−1 resulting in a sensitive, portable and integrated DNA analysis for a secondary metabolite.