• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.2
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• pp.132-136
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• 2004

The LAL (Limulus amebocyte lysate) test for the detection and quantification of endotoxin is based on the gelation reaction between endotoxin and LAL from a blood extract of Limulus polyphemus. The test is labor intensive, requiring dedicated personnel, a relatively long reaction time (approximately 1 h), relatively large volumes of samples and reagents and the detection of the end-point is rather subjective. To solve these problems, a miniaturized LOC (lab-on-a-chip) prototype, 62mm (L) ${\times}$ 18 mm (W), was fabricated using PDMS (polydimethylsiloxane) bonded to glass. Using this prototype, in which 2mm (W) ${\times}$ 44.3mm (L) ${\times}$ 100 $\mu\textrm{m}$ (D) microfluidic channel was constructed, turbidometric and chromogenic assay detection methods were compared, and the chromogenic method was found the most suitable for a small volume assay. In this assay, the kinetic-point method was more accurate than the end-point method. The PDMS chip thickness was found to be minimized to around 2 mm to allow sufficient light transmittance, which necessitated the use of a glass slide bonding for chip rigidity. Due to this miniaturization, the test time was reduced from 1 h to less than 10 min, and the sample volume could be reduced from 100 to ca. 4.4 ${\mu}$L. In summation, this study suggested that the LOC using the LAL test principle could be an alternative as a semi-automated and reliable method for the detection of endotoxin.

• Journal of the Korean Applied Science and Technology
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• v.28 no.1
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• pp.15-21
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• 2011

Threelectrodes systems were used in stripping voltammetry (SW) and cyclic voltammetry (CV) instead of the expensive platinum and Ag/AgCl reference electrodes. Moreover, the electrolyte solution was used with deep seawater, which can reduce water pollution, is more eco-friendly, and has a lower cost. The analytical optimum parameters measured via CV and SW and with working ranges were obtained from 10 to 80 ug/L using fluorine immobilized on a graphite pencil electrode (FE). Under the optimum conditions, the analytical detection limit of 6.30 ug/LAu was obtained. The results of the study can be applied to diagnostic assay for natural minerals and human finger tissue.

• Bulletin of the Korean Chemical Society
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• v.23 no.4
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• pp.599-603
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• 2002

A selective enzyme-linked immunosorbent assay (ELISA) for the insecticide isofenphos was developed. Three different analogues (haptens) of isofenphos were synthesized and were coupled to carrier proteins through the pesticide thiophosphate group t o use as immunogens or coating antigens. Rabbits were immunized with one of the haptens coupled to BSA for production of polyclonal antibodies and the sera were screened against each of the other two haptens coupled to ovalbumin (OVA). Using the sera of highest specificity, an antigen-coated ELISA was developed, which showed an I50 of 96 ng/mL with the detection limit of 2 ng/mL. The antibodies showed negligible cross-reactivity with other organophosphorus pesticides and the phenol metabolite of isofenphos, which makes the developed assay suitable for the selective detection of isofenphos. An antibody-coated ELISA was also developed, which showed an I50 of 580 ng/mL with a detection limit of 70 ng/mL.

• Bulletin of the Korean Chemical Society
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• v.23 no.10
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• pp.1399-1426
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• 2002

A competitive enzyme-linked immunosorbent assay (ELISA) was developed for the quantitative detection of the organophosphorus insecticide bromophos. Three bromophos analogues (haptens) were synthesized and were coupled to carrier proteins to use as immunogens or coating antigens. Rabbits were immunized with either one of two haptens coupled to bovine serum albumin (BSA) for production of polyclonal antibodies, and the sera were screened against one of the haptens coupled to ovalbumin (OVA). Using the serum with highest specificity and an enzyme tracer, an antibody-coated ELISA was developed, which showed an $IC_{50}$ of 40 ng/mL with a detection limit of 7 ng/mL. The antibodies in this assay showed negligible cross-reactivity with other organophosphorus pesticides except with the insecticides chlorpyrifos and fenitrothion.

• Bulletin of the Korean Chemical Society
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• v.17 no.11
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• pp.1018-1022
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• 1996

Adenosine deaminase (ADA) has been utilized as the label in devising a potentiometric homogeneous assay for riboflavin and riboflavin binding protein (RBP). The proposed homogeneous assay method employs an ADA-biotin conjugate as the signal generator and an avidin-riboflavin conjugate as the signal modulator in the solution phase. The catalytic activity of the ADA-biotin conjugate is inhibited in the presence of an excess amount of the avidin-riboflavin conjugate, and the observed inhibition is reversed in an amount proportional to the concentration of RBP added. When the analyte riboflavin is added to this mixture of ADA-biotin, avidin-riboflavin and RBP, the activity of the enzyme conjugate is re-inhibited in an amount proportional to the concentration of riboflavin. Since the enzyme label used in this system is ADA, an ammonia-producing enzyme, a potentiometric rather than photometric detection scheme is used to monitor the enzymatic activity in the assay.

• Bulletin of the Korean Chemical Society
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• v.22 no.4
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• pp.417-420
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• 2001

• Journal of the Korean Chemical Society
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• v.65 no.5
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• pp.397-400
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• 2021

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.2
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• pp.228-236
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• 2004

Two varieties of alfalfa (Medicago sativa L cv. Pioneer and Beaver) and timothy (Phleum pratense L cv. Climax and Joliette), grown at different locations in Saskatchewan (Canada), were cut at three stages [1=one week before commercial cut (early bud for alfalfa; joint for timothy); 2=at commercial cut (late bud for alfalfa; pre-bloom head for timothy); 3=one week after commercial cut (early bloom for alfalfa; full head for timothy)]. The energy values of forages were determined using three approaches, including chemical (NRC 2001 formula) and biological approaches (standard in vitro and in situ assay). The objectives of this study were to determine the effects of forage variety and stage of maturity on energy values under the climate conditions of western Canada, and to investigate relationship between chemical (NRC 2001 formula) approach and biological approaches (in vitro and in situ assay) on prediction of energy values. The results showed that, in general, forage species (alfalfa vs. timothy) and cutting stage had profound impacts, but the varieties within each species (Pioneer vs. Beaver in alfalfa; Climax vs. Joliette in timothy) had minimal effects on energy values. As forage maturity increased, the energy contents behaved in a quadratic fashion, increasing at stage 2 and then significantly decreasing at stage 3. However, the prediction methods-chemical approach (NRC 2001 formula) and biological approaches (in vitro and in situ assay) had great influences on energy values. The highest predicted energy values were found by using the in situ approach, the lowest prediction value by using the NRC 2001 formula, and the intermediate values by the in vitro approach. The in situ results may be most accurate because it is closest to simulate animal condition. The energy values measured by biological approaches are not predictable by the chemical approach in this study, indicating that a refinement is needed in accurately predicting energy values.

• Environmental Mutagens and Carcinogens
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• v.21 no.1
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• pp.34-43
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• 2001

Di-2-ethylhexyl phthalate (DEHP) is the most commonly used phthalate ester in polyvinyl chloride formulations including food packing and storage of human blood. DEHP is a well known as non-genotoxic carcinogen and endocrine disrupting chemical (EDC). DEHP have shown all negative results in ICH-guildeline recommended standard genotoxicity test battery. In this study, to assess the clastogenic and DNA damaging effect in human-derived tissue specific cells, DEHP was treated in human derived MCE-7 cells, HepG2 cells, LNCap cells, BeWo cells, MCE-10A cells, and female peripheral blood cells using micronucleus assay and in human breast carcinoma MCF-7 cells up to $1.28$\times$10^{-2}$ M using Comet assay. The in vitro micronucleus assay is a mutagenicity test system for the detection of chemicals which induce the formation of small membrane bound DNA fragment i.e. micronuclei in the cytoplasm of interphase cells, originated from clastogenic and/or aneugenic mechanism. The single cell gel electrophoresis assay (Comet assay) is used to detect DNA strand-breaks and alkaline labile site. In our results, DEHP increased significantly and/or dose-depentently and time-dependently micronucleus frequency at the 6 and 24 hr without metabolic activation system only in MCE-7 cells. DEHP treated with 2 hrs in MCF-7 cells using Comet assay induced DNA damage dose-depentantly.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.689-689
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• 2003