• Journal of Animal Science and Technology
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• 제57권4호
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• pp.13.1-13.5
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• 2015

As an alternative to radioimmunoassay a simple and highly sensitive enzyme immunoassay (EIA) was developed and validated for androstenedione quantification in plasma of Karan Fries bulls using second antibody coating technique. The wells of the microtitreplate were coated with affinity-purified goat immunoglobulin (antirabbit IgG) that binds the hormone specific antibody. The EIA was performed to analyze androstenedione directly in $40{\mu}l$ of bull plasma. The androstenedione standards ranged from 0.20 to 200 pg/$40{\mu}l$/well and the sensitivity of the assay was 5 pg/ml plasma. Serially diluted bull plasma containing high endogenous androstenedione showed good parallelism with bovine androstenedione standard curve. Intra- and inter-assay coefficients of variation (CV) were found to be 8 and 9%, respectively. Peripheral plasma androstenedione concentrations determined in young and adult bull samples ranged between 104-990 pg/ml and 184-2040 pg/ml, respectively.

• 대한수의학회지
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• 제46권3호
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• pp.291-294
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• 2006

$Parallux^{TM}$, a solid-phase fluorescence immunoassay(SPFIA) developed for antibiotics residue detection in milk, was applied for analysis of fish muscle. The recommended therapeutic dose of ampicillin(100 mg/kg body weight, withdrawal period 7 days) was orally administered to a group of 25 olive flounders(Paralichthys olivaceus) for consecutive five days. Muscle was sampled after drug treatment 1st, 2nd, 3rd, 4th and 5th day. The concentration of ampicillin in muscle, determined by SPFIA, was compared with that of internal standard(10 ppb as ampicillin). The absorbance ratio of sample to internal standard(Bs/Bo) was employed as an index to determine the muscle residue in olive flounder. To investigate the recovery rate, the standard solutions were added to muscle samples to give final concentrations in muscle of 10 and 50 ng/ml. The recovery rates of all spiked samples were > 89% of the spiked value. Ampicillin was detected in muscle of fishes treated until the 3rd day of withdrawal period. The present study showed that the SPFIA can be easily adopted in predicting tissue residues for ampicillin in farmed fishes.

• Bulletin of the Korean Chemical Society
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• 제24권1호
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• pp.70-74
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• 2003

The heterogeneous bioluminescence immunoassay for digoxin was developed using photoprotein, native aequorin as a label and the site-directed immobilization technique based on avidin/biotin interaction. Aequorin is a bioluminescence protein, originally isolated from the jellyfish Aequoria Victoria and an attractive label in analytical applications because of sensitive detection due to virtually no background bioluminescent signal. Digoxin is a cardioactive drug, and its therapeutic level in serum is at low concentration with very narrow therapeutic index. The aequorin-digoxigenin conjugates were synthesized by the N-hydroxysuccinimide ester method and characterized in terms of bioluminescent residual activity. The resulting dose-response curve shows that the detection limit is $1.0\;{\times}\;10^{-10}\;M$ and a dynamic range is three orders of magnitude, which was obtained by $1.0\;{times}\;10^{-10}\;M$ conjugate and 0.9 μg/mL anti-digoxin antibody. Three structurally similar molecules to digoxin were examined for their cross-reactivity. None of these three compounds showed any crossreactivity with digoxin antibody employed in this study. Standard amounts of digoxin corresponding to the therapeutic range were spiked into the each serum solution. Study of the serum matrix effect indicated that correlation coefficient shows good agreement between luminescence light intensity between in buffer and in serum.

• 대한임상검사과학회지
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• 제42권1호
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• pp.32-37
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• 2010

The author was performed to investigation of current status of prevalence for anti-hepatitis C virus (HCV) among the health-checkup adults in Jeonbuk province. A toal of 1,553 (male 1,046, female 507) serum samples were diagnosed by 3rd generation enzyme immunoassay (EIA) for anti-HCV. Total prevalence of anti-HCV was 0.9%, and prevalence of male and female were 0.8% and 1.2%, respectively. The prevalence of female was higher than male. According to ages group, prevalence of anti-HCV was highest in 60 age group, but it was not found in 20 age group. 14 samples with anti-HCV positive were diagnosed by EIA for hepatitis B virus surface antigen (HBs Ag), by chemiluminescence immunoassay (CLIA) for serum albumin, alanine transaminase (ALT) and asparagine transaminase (AST). Positive for HBs Ag was not found. The mean of serum albumin levels was 4.5 g/dL, and mean of ALT and AST were 34.3 IU and 31.9 IU, respectively. Through this study, I know that the prevalence of anti-HCV among adults in Jeonbuk, and suggest that the positive of anti-HCV persons who have lower serum albumin, normal to mild elevations in serum enzymes are chronic hepatitis.

• 대한수의학회지
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• 제33권4호
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• pp.611-615
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• 1993

젖소에서 progesterone과 testosterone의 측정을 위하여 solid phase 효소면역분석법(酵素免疫分析法)를 개발하였다. 1차 항체로서 $11{\alpha}$-hemisuccinate-progesterone bovine serum albumin과 4-androsten-$17\;{\beta}$-ol-3-one-carboxymethyloxime bovine serum albumin에 대한 토끼 혈청(血淸)을 각각 항혈청(抗血淸)으로 사용하였고, 2차 항체로서 면약 IgG를 사용하였다. Conjugates로서는 각각 progesterone-$11\;{\alpha}$-hydroxy-hemisuccinate-horseradish peroxidase와 4-androsten-$17\;{\beta}$-ol-3-hemisuccinate-horseradish peroxidase를 사용하였다. Progesterone과 testosterone에 대한 최저측정치(最低測定値)는 각각 6.7 pg/well과 1.0 pg/well이었다.

• Journal of Animal Science and Technology
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• 제64권4호
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• pp.621-639
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• 2022

Cortisol and corticosterone, hormones traditionally considered biomarkers of stress, can be measured in fluid biomatrices (e.g., blood, saliva) from live animals to evaluate conditions at sampling time, or in solid biomatrices (e.g., hair, feather) from live or dead animals to obtain information regarding long-term changes. Using these biomarkers to evaluate physiological stress responses in domestic animals may be challenging due to the diverse characteristics of biomatrices for potential measurement. Ideally, a single measurement from the biomatrix should be sufficient for evaluating chronic stress. The availability of appropriate and cost-effective immunoassay methods for detecting the biomarkers should also be considered. This review discusses the strengths and limitations of different biomatrices with regard to ensuring the highest possible reliability for chronic stress evaluation. Overall, solid biomatrices require less frequent sampling than other biomatrices, resulting in greater time- and cost-effectiveness, greater ease of use, and fewer errors. The multiplex immunoassay can be used to analyze interactions and correlations between cortisol and other stress biomarkers in the same biomatrix. In light of the lack of information regarding appropriate biomatrices for measuring chronic stress, this review may help investigators set experimental conditions or design biological research.

• 대한의생명과학회지
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• 제2권1호
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• pp.41-48
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• 1996

환자의 복수와 늑막액으로부터 p-diazonium phenylphosphorylcholine(DPPC) coupled Separose-4B affinity chromatography와 hydroxylapatite chromatography를 실시하여 C-reactive protein (CRP)를 분리, 정제하였다. 정제된 CRP를 토끼에게 면역화하여 항혈청을 얻고 affinity chromatography를 하여 면역항체(IgG)를 분리하였다. 분리된 면역항체를 미립자에 감작시킨 후 미립자 응집반응에 의하여 3분내에 CRP를 측정할 수 있는 간이 면역 측정법을 개발하였다. 본 연구에서 개발된 CRP측정법의 검출범위는 0.5~20mg/㎗이며, 임상 시험 결과 0.7~2.9mg/㎗에서는 강한 응집 반응을, 5.O~l3.2mg/㎗에서는 약한 응집반응을 보였고 28mg/dl이상에서는 항원 과잉으로 인한(zone of Ag excess phenomenon) 위음성을 나타냈다. 74명의 환자 혈청을 대상으로 CRP의 농도를 조사한 결과 평균치는 3.8mg/dl이었으며 대부분의 환자에서는 10mg/dl 이하의 농도로 존재하였다. 그러므로 1차 판정시 음성을 나타낸 시료라도 혈청을 5~10배정도 희석하여 재분석한다면 오차없이 CRP 를 검출할 수 있었다. 환자 혈청을 검체로 하여 본 연구에서 개발한 면역측정법과 현재 수입 시판 중인 프랑스의 B사 제품과 일본의 I사 제품을 비교한 결과 좋은 상관관계를 보였다. 이와 같은 평가 분석을 통하여 볼 때 본 연구에서 개발한 간이 면역 측정법은 사용이 비교적 간편하며 신빙성이 있어 CRP를 스크리닝하는데 효과적임을 알 수 있었다.

• 한국수산과학회지
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• 제48권2호
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• pp.158-167
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• 2015

A lateral flow immunoassay kit based on antigen-antibody interactions was developed to detect residues of beta-lactams, quinolones, tetracyclines, and sulfonamides in farmed fish. Group-specific antibodies showing cross-reactivity with other antibiotics in the same group were produced in rabbits. The rabbits were immunized eight times to obtain the maximum titers. Antibodies were extracted from the antisera collected from the immunized rabbits and produced group-specific reactions with antibiotics from the four groups. A kit was prepared that optimize conditions for the antigen-antibody reaction, using colloidal gold conjugated antibodies, and was designed to detect the four groups of antibiotics simultaneously. The kit enabled the detection of antibiotics in the four groups at below maximum residue limits (MRLs), which were $200{\mu}g/kg$ for tetracyclines, $100{\mu}g/kg$ for sulfonamides, $50{\mu}g/kg$ for beta-lactams, and $100{\mu}g/kg$ for quinolones. The cross-reactivity of the antibodies ranged from 10-80% for the sulfonamides, 20-100% for tetracyclines, 38-100% for quinolones, and 20-100% for the beta-lactams, confirming that the antibodies were group specific. The test kit was used 30 times to examine spiked antibiotics at the limits of detection (LODs) and all produced positive results, indicating high sensitivity. The LODs for the assay ranged from 4-20 ng/mL for beta-lactams, 25-50 ng/mL for sulfonamides, 20-100 ng/mL for tetracyclines, and 30-80 ng/mL for quinolones, and there were no false negative reactions at above these LODs. In addition, all of the LODs of the developed kit were correlated with high-performance liquid chromatography (HPLC) data. Our lateral flow immunoassay kit can simultaneously detect antibiotic residues from a large number of fish samples rapidly, strengthening the safety of domestic farmed and imported fish.

• 한국식품과학회지
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• 제41권1호
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• pp.111-115
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• 2009

Enzyme-linked fluorescent immunoassay(ELFA)를 이용하여 가공식품 중 땅콩을 분석하고, 그 결과가 표시내용과 일치하는지를 조사하였다. 땅콩으로부터 분리한 조단백질(CPP)을 토끼에게 면역하여 생산한 특이항체 및 horseradish peroxidase-항체 복합물을 이용하여 sandwich ELFA를 확립하였다. 특이항체의 교차반응은, CPP, 땅콩, 아몬드, 콩, 호도에 대하여 각각 100, 9.8, $1.1{\times}10^{-2},\;4.4{\times}10^{-3}$, 0%로 나타났다. 시료로는 비스킷, 스낵, 초콜릿 등 19점을 사용하였다. 이들의 분석결과, 7점의 시료에서 땅콩이 검출 되었으며, 이 중 땅콩의 함유가 표시된 제품은 5점이었고 그렇지 않은 제품은 2점이었다. 이들 2점 중 하나는 아몬드를 함유하고 있다고 표시된 비스킷으로 분석 상 미량($2.1{\times}10^{-3}%$)의 땅콩이 검출되었는데, 이는 교차반응 때문으로 추측된다. 나머지 1점은 대두의 함유가 표시된 스낵으로 분석 상 $9.8{\times}10^{-2}%$의 땅콩이 검출되었다. 이는 대두의 매우 낮은 교차반응율을 감안하면 제조공정상 땅콩의 교차오염 때문으로 추측된다. 이와 같이 ELFA에 의하여 가공식품 중 땅콩의 검출이 가능하였고, 현재 유통 되는 일부 가공식품 중 알레르겐의 표시를 정확히 할 필요가 있다고 생각한다.

• 한국자기학회:학술대회 개요집
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• 한국자기학회 2007년도 The 1st International Symposium on Advanced Magnetic Materials
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• pp.93.1-93.1
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• 2007