Ok-Jae Sohn

Sequential injection analysis system for on-line monitoring of l-cysteine concentration in biological processes

A sequential injection analysis (SIA) system was developed to monitor the concentration of l-cysteine in biological processes on-line. It is based on the redox reaction of l-cysteine with iron(III) in the presence of 1,10-phenanthroline (phen) and the detection of the red-iron(II)-phen complex with a spectrophotometry. The system was fully automated using software written in the LabVIEWtrade mark development environment. A number of system variables such as the flow rate of the carrier buffer solution, the volume ratio of the sample to the reagents, and the reaction coil length, etc., were evaluated to increase the sensitivity and performance of the SIA system. Under partially optimized operating conditions the performance of the SIA system was linear up to a concentration of l-cysteine of 1mM (R(2)=0.998) with a detection limit of 0.005mM and a sample frequency of 15hr(-1). The SIA system was employed to monitor the concentration of l-cysteine on-line in a continuously stirred reactor and a fermentation process of Saccharomyces cerevisiae. The on-line monitored data were in good agreement with the off-line data measured by a HPLC with a fluorescence detector (n=15, R(2)=09899).

Application of principal component analysis and self-organizing map to the analysis of 2D fluorescence spectra and the monitoring of fermentation processes

Abstract2D fluorescence sensors produce a great deal of spectral data during fermentation processes, which can be analyzed using a variety of statistical techniques. Principal component analysis (PCA) and a self-organizing map (SOM) were used to analyze these 2D fluorescence spectra and to extract useful information from them. PCA resulted in scores and loadings that were visualized in the score-loading plots and used to monitor various fermentation processes with recombinantEscherichia coli andSaccharomyces cerevisiae. The SOM was found to be a useful and interpretative method of classifying the entire gamut of 2D fluorescence spectra and of selecting some significant combinations of excitation and emission wavelengths. The results, including the normalized weights and variances, indicated that the SOM network is capable of being used to interpret the fermentation processes monitored by a 2D fluorescence sensor.

Flow injection analysis system for monitoring of succinic acid in biotechnological processes

In this study, a flow injection analysis (FIA) system using a cartridge of immobilized isocitrate lyase (ICL) and isocitrate dehydrogenase (ICDH) was developed to monitor the concentrations of succinic acid in biotechnological processes. The ICL and ICDH immobilized on VA-Epoxy Biosynth E3-carrier had a good operational lifetime (up to 24h) and storage stability (up to 30 days). The FIA system with the immobilized ICL/ICDH cartridge was characterized with respect to the factors affecting the activity of the immobilized enzymes, such as pH of carrier solution, temperature, sample matrix, etc. Optimal pH value of the immobilized enzymes was slightly shifted in the alkaline range, i.e. 9.0. Some components such as 10gl(-1) lactose, 3gl(-1) malate and 3gl(-1) oxaloacetate in sample solution had significant activating effects (more than 10%) on the response of the FIA system. But the activity of the immobilized ICL and ICDH was not largely influenced by some components like imidazole (1mM), sodium azide (10mM) and semicarbazide (2gl(-1)) added to carrier buffer solution. The FIA system with an enzyme cartridge was applied to on-line monitor the concentrations of succinic acid in a continuously stirred reactor and a fermentation process of immobilized Escherichia coli, and showed good sensitivity and reliability of the FIA system developed in this work.

Development of optical immunosensors and their application to the analysis of human bone morphogenetic protein-7 (BMP-7)

In this study, a few optical immunosensors were developed to determine the concentration of BMP-7. Hydrophilic CdSe/ZnS quantum dots (QDs) were synthesized and conjugated to the antibody of BMP-7 (BMP-7Ab). The QDconjugated BMP-7Ab was used as a fluorescence probe at excitation and emission wavelengths of 470 nm and 585 nm, respectively. It was immobilized either on the bottom of the well of a 96-well microtiter plate or on the tip of an optical fiber. Two immunoassays, i.e. the direct and sandwich assays, were studied for their sensitivity. The sensitivity of the direct immunoassay was 1296.21, compared to 384.69 for the sandwich assay. The linear detection range was 0.0-1.0 ng/mL for both assays. Based on the results of the microtiter plate technique, the direct assay technique was used for the development of an optical fiber immunosensor. The optical fiber immunosensor has a linear detection range between 0.0 and 10.0 ng/mL with a detection limit of 0.413 ng/mL. The optical fiber immunosensor was applied to the sequential injection analysis for the automatic determination of BMP-7.

Optical sensing techniques for simultaneous detection of nanoparticles and microorganisms in water

An optical sensor was developed to detect nanoparticles, turbid materials and microorganisms in water simultaneously. Three different light sources like UV-LED, NIR-LED and laser diode have been employed to develop the optical sensor based on the scattering light and fluorescence light. The sensor system has high selectivity and sensitivity, that it can be used to monitor the quality of drinking water.

Fiber-optic biosensor for analysis of glucose and lactate in blood samples

Optical-fiber sensors have been developed to determine the concentrations of glucose and lactic acid in blood samples. Fluorescence dye [tris(2,2`-biphenyridine)-ruthenium(II)-chloride (RuBPY)] was entrapped by using a silicon to the unclad tip of a glass optic fiber. Enzymes like glucose oxidase (GOD) and lactate oxidase (LOD) have been immobilized by acrylamide resin adhesive, adsorption with zeolite or covalent bonding with aminopropyl-triethoxysilan. The fiber-optic glucose/lactate sensor was then used to analyze the concentrations of glucose and lactate in blood samples. The results were compared with the results of HPLC analysis and their difference was in error by less then 5 %.