Wen-Yih Chen

The investigation of recognition interaction between phenylboronate monolayer and glycated hemoglobin using surface plasmon resonance

Glycated hemoglobin (HbA1c) is formed by a nonenzymatic reaction of glucose with the N-terminal valine of adult hemoglobins beta-chain. The amount of HbA1c reflects the average concentration of glucose variation level over the preceding 2 to 3 months. Because the boronate has antibody mimicking for HbA1c, often it is used to detect HbA1c. However, factors such as the ratio of the phenylboronic acid derivatives and diol composition, the pH of the solution, and the stereostructure of phenylboronic acid derivatives could influence the interactions between phenylboronic acid derivatives and diol composition. In this study, the factors were evaluated using surface plasmon resonance (SPR). The results show that pH value is an important factor affecting HbA1c and phenylboronic acid to form the complex and Lewis bases. This could change the stereostructure of phenylboronic acid to form B(OH)(3) for binding with saccharine easily. In addition, linear response appeared in HbA1c in the range of 0.43 to 3.49 mug/ml, and the detection limit was 0.01 microg/ml. The results also demonstrated that an SPR biosensor can be used as a sensitive technique for improving the accuracy and correctness of HbA1c measurement.

A Guided Mode Resonance Aptasensor for Thrombin Detection

Recent developments in aptamers have led to their widespread use in analytical and diagnostic applications, particularly for biosensing. Previous studies have combined aptamers as ligands with various sensors for numerous applications. However, merging the aptamer developments with guided mode resonance (GMR) devices has not been attempted. This study reports an aptasensor based home built GMR device. The 29-mer thrombin aptamer was immobilized on the surface of a GMR device as a recognizing ligand for thrombin detection. The sensitivity reported in this first trial study is 0.04 nm/μM for thrombin detection in the concentration range from 0.25 to 1 μM and the limit of detection (LOD) is 0.19 μM. Furthermore, the binding affinity constant (Ka) measured is in the range of 106 M−1. The investigation has demonstrated that such a GMR aptasensor has the required sensitivity for the real time, label-free, in situ detection of thrombin and provides kinetic information related to the binding.

Real-time monitoring DNA hybridization by guided resonant mode biosensor

In this paper, a label free approach of the DNA hybridization detection by the guided-mode resonance (GMR) spectroscopy sensor is presented. The GMR biosensor can easily detect any slight variation of the DNA hybridization by the resonant wavelength peak shift. The corresponding peak wavelength shifts of the capture DNA, the target DNA and the probe DNA serially hybridized reactions from GMR are measured, respectively. Furthermore, the stability and reliability of the GMR biosensor for nucleic acid hybridization have also been examined. The result obviously reveals the feasibility of implementation of the GMR-based DNA microarray for detecting on the dynamics of DNA hybridization.

Real-time monitoring on peptide synthesis by GMR biosensors

In this work, a real-time monitoring system has been developed. The micro-optic device applying the guided-mode resonance (GMR) effect has been designed and fabricated. By combining a simple and low cost microfluidic channel, the GMR measurement for detecting peptide synthesis reaction has been explored. It is able to determine whether or not the chemical reaction reacts and how far the chemical reaction arrived. In addition, the kinetics analysis for the peptide synthesis has also been discussed. It is successfully demonstrated that the GMR device is capable of monitoring the process of peptide synthesis in real time.

Isothermal titration calorimetry for characterization of recombinant proteins

Isothermal titration calorimetry is widely used to measure the affinities and enthalpies of interaction between proteins and/or small molecules. The quantitative nature of the technique is especially useful in the characterization of recombinant proteins while determining the fraction of protein capable of binding a specific ligand and thus the protein purity. The revealed thermodynamic information sheds light on the binding mechanism, important for the targeted drug design of the biologics. Here we show examples how, together with the thermal shift assay, combination of both techniques enables characterization of protein stability and ligand binding. Furthermore, the binding-linked reactions that strongly affect the observed thermodynamic parameters and must be dissected to obtain the intrinsic parameters that are necessary for the structure-based rational drug design are being demonstrated using inhibitors of Hsp90, an anticancer target protein.