Seung Yeon Song

Multienzyme-modified biosensing surface for the electrochemical analysis of aspartate transaminase and alanine transaminase in human plasma

AbstractWe investigated the electrochemical detection of aspartate transaminase (AST) and alanine transaminase (ALT) by using a multienzyme-modified electrode surface. Determination of the activities of transaminases in human serum is clinically significant because their concentrations and ratios indicate the presence of hepatic diseases or myocardial dysfunction. For electrochemical detection of AST and ALT, enzymes that participate in the reaction mechanism of AST and ALT, such as pyruvate oxidase (POX) and oxaloacetate decarboxylase, were immobilized on an electrode surface by using an amine-reactive self-assembled monolayer and a homobifunctional cross-linker. In the presence of suitable substrates such as l-aspartate (l-alanine) and α-ketoglutarate, AST and ALT generate pyruvate as an enzymatic end product. To determine the activities of AST and ALT, electroanalyses of pyruvate were conducted using a POX and ferrocenemethanol electron shuttle. Anodically generated oxidative currents from multienzyme-mediated reactions were correlated to AST and ALT levels in human plasma. On the basis of the electrochemical analysis, we obtained calibration results for AST and ALT concentrations from 7.5 to 720 units/L in human plasma-based samples, covering the required clinical detection range. FigurePOX-OAC calatytic cycles for AST and ALT analysis

Chip-based cartilage oligomeric matrix protein detection in serum and synovial fluid for osteoarthritis diagnosis

We have developed a method to detect cartilage oligomeric matrix protein (COMP) as a specific biomarker of osteoarthritis (OA). In pathological conditions of the cartilage, COMP is released first into the synovial fluid (SF) and from there into the blood. Thus, measurement of COMP in the blood and SF facilitates OA diagnosis. To determine COMP, we developed a fluoro-microbead guiding chip (FMGC)-based immunoassay. The FMGC has four immunoreactive regions, each with five patterns, to allow multiple assays. A COMP-specific capture antibody was immobilized to the FMGC surface to create a self-assembled interfacial layer. SF or serum samples from patients with OA possessing the target COMP were applied to the COMP-sensing monolayer. To generate binding signal, COMP detection antibody-conjugated fluoro-microbeads were applied and the numbers of fluoro-microbeads bound specifically were counted to determine COMP concentrations. This FMGC-based immunoassay clearly distinguished immunospecific from nonspecific binding by comparing optical signals from inside and outside of the patterns. The optical signals showed linear correlations with serum and SF COMP concentrations. Optical detection and quantification of COMP using fluorescence microscopy correlated well with results from commercial enzyme-linked immunosorbent assay (ELISA). This FMGC-based immunoassay offers a new approach for detecting a clinically relevant biomarker for OA in human blood and SF.

Development of a Wireless Love Wave Biosensor Platform for Multi-functional Detection

A Love wave-based biosensor with a 440 MHz operating frequency was developed for simultaneous detection of two different concentrations of anti-dinitrophenyl-keyhole limpet hemocyanin (Anti-DNP-KLH) rabbit immunoglobulin G (IgG) in a single sensor. The sensor was composed of surface acoustic wave (SAW) reflective delay lines built from interdigital transducer (IDT) and several reflectors, a poly(methyl methacrylate) (PMMA) waveguide layer, and two sensitive films. To extract optimal device parameters, coupling of mode (COM) modeling was carried. According to the device parameters determined, the Love wave biosensor was fabricated and then wirelessly characterized by a network analyzer. Binding of anti-DNP IgG to DNP induced a change in the time positions of the original reflection peaks mainly due to the mass loading effect. The measured time positions were matched well with the predicted values from COM modeling. The sensitivities evaluated from the first and second sensitive films were 167.9 and 44.8 degµg-1ml-1, respectively.

A wireless love wave biosensor platform for simultaneous detections of two different biomolecules

Love wave biosensor operating at 440 MHz was developed for simultaneous detection of anti-DNP-KLH IgG in a single sensor. The developed sensor was composed of surface acoustic wave (SAW) reflective delay lines structured by interdigital transducer (IDT) and shorted grating reflectors, a waveguide layer, and two sensitive films. Two mass loading effects caused by two different IgG concentrations were simultaneously measured in a single sensor. The evaluated sensitivities from the first and second sensitive film were 167.9 °/µg/ml and 44.8 °/µg/ml, respectively.