Byung-Wook Park

Recent progress in bio-sensing techniques with encapsulated enzymes

Biosensors with encapsulated enzymes have advantages of high substrate selectivity and sustained enzyme activity. Enzymes are encapsulated in various materials, such as liposome, polymer, and sol-gel or hydro-gel, depending on sensing conditions. By stabilizing the enzymes via encapsulation with new methods and materials the enzyme activity may be maintained for a longer time, and even the selectivity can possibly be enhanced. In general increased mass transfer limitation seems to be the major challenge to investigate more. Novel materials, encapsulation techniques, and sensing mechanisms have been studied intensively for encapsulated enzyme biosensors. In this review, we focus on the recent progress in encapsulated enzyme biosensors published in peer-reviewed journals over the last 10 years. The articles are categorized and reviewed in four groups based on encapsulation techniques incorporating with liposome, polymer, sol-gel or hydro-gel, and peptide. Research articles that are considered critical and noticeable are selected and compared according to these categorizations. Based on these article analyses, we suggested future direction in the encapsulated enzyme biosensor research.

A novel glucose biosensor using bi-enzyme incorporated with peptide nanotubes.

A novel amperometric glucose biosensor was developed using the bio-inspired peptide nanotube (PNT) as an encapsulation template for enzymes. Horseradish peroxidase (HRP) was encapsulated by the PNT and glucose oxidase (GO(x)) was co-immobilized with the PNT on a gold nanoparticle (AuNP)-modified electrode. A binary SAM of 3-mercaptopropionic acid (MPA) and 1-tetradecanethiol (TDT) was formed on the surface of the electrode to immobilize the PNT and GO(x). The resulting electrode appeared to provide the enzymes with a biocompatible nanoenvironment as it sustained the enhanced enzyme activity for an extended time and promoted possible direct electron transfer through the PNT to the electrode. Performance of the biosensor was evaluated in terms of its detection limit, sensitivity, pH, response time, selectivity, reproducibility, and stability in a lab setting. In addition the sensor was tested for real samples. The composite of AuNP-SAM-PNT/HRP-GO(x) to fabricate a sensor electrode in this study exhibited a linear response with glucose in the concentration range of 0.5-2.4mM with a R(2)-value of 0.994. A maximum sensitivity of 0.3 mA M(-1)and reproducibility (RSD) of 1.95% were demonstrated. The PNT-encapsulated enzyme showed its retention of >85% of the initial current response after one month of storage.

Enzyme activity assay for horseradish peroxidase encapsulated in peptide nanotubes.

Encapsulation of horseradish peroxidase (HRP) inside a peptide nanotube (PNT) was demonstrated and its activity was measured. Enzyme assay verified that 0.16 μg of the enzymes were encapsulated in 1mg of PNTs. The encapsulation was also verified with TEM, UV-vis spectroscopy, and FTIR. The activity of the encapsulated HRP was examined for thermal stability, long-term storage stability, and resistance to a denaturant. They showed good storage stability, retaining its activity up to 90%, while the free HRP lost 50% of its activity over the course of 18 days. At 55 °C, the encapsulated HRP activity remained 20% higher than that of the free HRP. With the denaturant, guanidinium hydrochloride (GdmHCl), the encapsulated HRP activity was maintained around 10% higher than the free HRP. This result proves that the encapsulation of HRP inside the PNT may be an effective way to keep the enzyme activity stable in various environments.

Development of a highly specific amine-terminated aptamer functionalized surface plasmon resonance biosensor for blood protein detection

This paper presents a generally applicable approach for the highly specific detection of blood proteins. Thrombin and thrombin-binding aptamers are chosen for demonstration purposes. The sensor was prepared by immobilizing amine-terminated aptamers onto a gold modified surface using a two-step self-assembled monolayer (SAM) immobilization technique and the physical detection is performed using Surface Plasmon Resonance (SPR). The developed sensor has an optimal detectable range of 5–1000 nM and the results show the sensor has good reversibility, sensitivity and selectivity. Furthermore, the sensor shows the potential of being improved and standardized for direct detection of other blood proteins for clinical applications.