• 한국연초학회지
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• 제19권1호
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• pp.5-10
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• 1997

Three strains of W isolated from tobacco, tomato and Pepper plants in Korea were characterized based on biological response, serological relationship, and peptide mapping of the capsid Proteins. The strains designated as TMV-common, TMV-Pepper, and TMV-tomato could be distinguishable by different visual symptoms on 3 varieties of tobacco, one variety of tomato and Pepper for each among 27 plant specieces. Serological relationships were examined by agar gel double diffusion test. Only traceable or weak reaction was observed in the incompatible antigen-antibody combinations. The Pepper strain, however, showed trace in reaction with other two antisera. Peptide maps of the capsid proteins digested by V8 protease or by trypsin were also distinguishable, suggesting differences in composition and/or sequence of the amino acids among the strains.

• 한국가금학회지
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• 제41권1호
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• pp.15-20
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• 2014

본 연구는 가금에서 호흡기 질환, 심낭수종, 봉입체 간염, 산란저하증 등을 유발하는 가금 아데노바이러스의 fiber 2 유전자에 대한 특이 난황 항체를 개발하고자 실험을 실시하였다. Fiber 2 유전자를 클로닝한 뒤, 대장균 발현 시스템을 이용하여 약 22 kDa의 재조합 fiber 2 단백질을 생산하였다. 이를 산란계에 3주 간격으로 총 4회 면역하여 혈청 및 난황 내 항체가를 측정한 결과, 면역 후 12주경에 항체가가 최고치에 달하였으며, 산란계로부터 획득한 난황 항체를 이용한 Western blot analysis 결과, 가금 아데노바이러스 내 fiber 2 단백과 특이적으로 반응한다는 것을 규명하였다. 결론적으로, 가금 아데노바이러스 fiber 2에 특이적인 난황 항체 생산에 성공하였으며, 이러한 특이적인 난황 항체는 가금 아데노바이러스로 인한 질병의 예방 및 치료에 활용 가능할 것으로 사료된다.

• Journal of Veterinary Science
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• 제22권4호
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• pp.45.1-45.13
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• 2021

Background: Peste des petits ruminants (PPR) is an infectious disease caused by the peste des petits ruminants virus (PPRV) that mainly produces respiratory symptoms in affected animals, resulting in great losses in the world's agriculture industry every year. Single-domain variable heavy chain (VHH) antibody fragments, also referred to as nanobodies, have high expression yields and other advantages including ease of purification and high solubility. Objectives: The purpose of this study is to obtain a single-domain antibody with good reactivity and high specificity against PPRV. Methods: A VHH cDNA library was established by immunizing camels with PPRV vaccine, and the capacity and diversity of the library were examined. Four PPRV VHHs were selected, and the biological activity and antigen-binding capacity of the four VHHs were identified by western blot, indirect immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) analyses. ELISA was used to identify whether the four VHHs were specific for PPRV, and VHH neutralization tests were carried out. ELISA and western blot analyses were used to identify which PPRV protein was targeted by VHH2. Results: The PPRV cDNA library was constructed successfully. The library capacity was greater than 2.0 × 10⁶ cfu/mL, and the inserted fragment size was approximately 400 bp to 2000 bp. The average length of the cDNA library fragment was about 1000 bp, and the recombination rate was approximately 100%. Four single-domain antibody sequences were selected, and proteins expressed in the supernatant were obtained. The four VHHs were shown to have biological activity, close affinity to PPRV, and no cross-reaction with common sheep diseases. All four VHHs had neutralization activity, and VHH2 was specific to the PPRV M protein. Conclusions: The results of this preliminary research of PPRV VHHs showed that four screened VHH antibodies could be useful in future applications. This study provided new materials for inclusion in PPRV research.

• 생약학회지
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• 제50권3호
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• pp.175-184
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• 2019

Mast cells are crucial as effector cells in the immediate-type allergic reaction. Lentinus edodes has been the popular edible mushroom in oriental countries and reported to have immunomodulatory, anti-tumor, anti-atherogenic, anti-viral, and anti-allergic activities. However, the roles of L. edodes in mast cell-mediated anaphylactic reaction have not been fully elucidated. In this research, we have demonstrated the effects of the methanol extract of L. edodes (MELE) on mast cell-mediated anaphylaxis-like and anaphylactic reactions. MELE suppressed systemic anaphylaxis-like reaction, plasma histamine levels, and ear swelling response in mice treated with compound 48/80. MELE also suppressed passive systemic and cutaneous anaphylaxis mediated by anti-dinitrophenyl IgE. In accordance with these findings, MELE dose-dependently decreased histamine release from RPMC evoked by compound 48/80 or the antigen-antibody reaction. To clarify the mechanism of degranulation system, intracellular cAMP levels as well as calcium influx in RPMC was evaluated. In compound 48/80-treated RPMC, MELE blocked calcium uptake into the cells. In addition, MELE elevated the intracellular cAMP content and significantly attenuated compound 48/80-induced cAMP reduction in RPMC. Taken together, we propose the clinical use of MELE in mast cell-mediated immediate-type allergic diseases.

• 한국어병학회지
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• 제12권1호
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• pp.24-31
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• 1999

어류의 edwardsiellosis를 진단하기 위한 solid phase ELISA법에 대한 최적화 분석을 실시하였다. 부분 정제한 E. tarda Edk-2에 대한 토끼 항혈청을 sodium bicarbonate 완충용액에 $50{\mu}g/ml$ 농도로 희석하여 overnight 반응시켜 주었을 때 polystyrene bead 표면의 항체 immobilization이 최적화되었다. $50{\mu}g/ml$의 biotin 표지화 항체와 1:2000으로 희석된 extravidin-peroxidase를 차례로 처리하였을 때 최적의 반응을 나타내었으며 이렇게 최적화시킨 정성적 solid phase ELISA법은 EDTA 추출법으로 조제된 항원에 대해서는 $1{\times}10^5$ cells/ml, 열 추출법으로 조제된 항원에 대해서는 $1{\times}10^5$ cells/ml의 검출한계를 나타내었다. E. tarda Edk-2에 대한 토끼 항혈청을 이용한 본 연구의 solid phase ELISA법은 우리나라 양어장의 넙치 병어로부터 분리한 여러 한국형 E. tarda 균주와도 높은 교차반응을 나타내었다. 이러한 것은 본 기법을 다른 지역에서 분리한 여러 strain의 E. tarda에 대한 진단을 위하여 본 실험의 항혈청을 사용하여 가능하다는 것을 보여 주었다.

• Parasites, Hosts and Diseases
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• 제22권2호
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• pp.223-228
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• 1984

외과적으로 충체를 적출하여 진단한 스파르가눔증 환자 7명으로부터 혈청을 수집하고 혈청내 스파르가눔 특이 IgG 항체가를 호소면역 측정 법으로 측정하였다. 스파르가눔 항원은 유혈목이에 자쳔 감염된 충체를 갈아 생리 식염수로 추출한 것으로 단백질 함량은 Smgyxnl이었다. 효소면역측정법은 McLaren 등(1978)의 방법에 따라 실시하였다. 스파르가눔중 환자 혈청이 폐흡충 항원 또는 유구낭미충 항원에 교차반응이 있는지 여부도 측정하였다. 또한 스파르가눔 항원에 대해서 정상인, 케흡충중 환자, 간홉충 감염자, 유구낭미충증 환자와 무구조충 감염자 등 모두 71명에 대해서도 교차반응을 나타내는지 를 관찰하고자 효소면역 측정 법을 실시하였다. 그 결과를 요약하면 다음과 같다. 1. 스파르가늠증 환자 7명중 석회화한 스파르가눔에 감염되어 있었고 수술후 1개월에 혈청을 수집한 1례를 제외하면 모두 홉광도 0.3 이상을 나타내었다. 양성판정기준을 흠광도 0.25로 하였을 때 효소면역 측정 법의 민감도(sensitivity)는 85.7이 었다. 2. 기타 기생충 감염자와 정상인 71명중에서 스파르가눔 항원에 대한 특이항체 양성자는 무구조충감염자 3명으로 특이도(specificity)는 97.5%이었다. 3. 스파르가눔중 환자 혈청은 폐흡충 항원 및 유구낭미충 항원에 대해서 항체가는 낮은 값을 보이고 있어 교차 반응은 발견할 수 없었다. 4. 중추신경계를 침범한 스파르가눔중 환자 2례에서 뇌척수액을 희석하지 않은 상태로 효소면역 측정 법을 실시하였던 바 1례는 폐흡충 및 유구낭미충 항원에 대해 교차반응 없이 스파르가눔항원에 대해 높은 항체가를 보인데 비해 다른 1례에서는 스파르가눔 항원 및 유구낭미충 항원에 대해 비슷한 항체가를 나타내고 있었다. 이상의 결과에서 효소면역 측정 법을 이용하여 스파르가눔중을 혈청학적으로 진단하는 경우 매우 특이하고 민감한 결과를 얻을 수 있을 것으로 생각하였다. 스파르가눔중의 수술전 진단이 실제로 어려운 것이 현실이므로 혈청학적 잖단은 역학적 유해조사나 형태학적으로 감별이 어려운 병리소견을 보인 경우 등에서 보조적인 진단법으로 이용할 수 있을 것으로 생각한다.

• Parasites, Hosts and Diseases
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• 제36권4호
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• pp.269-275
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• 1998

톡소포자충과 네오포자충 (Neospora caninum)의 충체 항원으로 톡소포자충 양성 혈청 및 톡소포자충증 환자의 혈청과 ELISA, western blot 및 면역형광법을 실시하였다. ELISA에서는 172명의 톡소포자충 양성 혈청에서 12명 （6.7％）이 두 항원에 모두 반응하였으며, 톡소포자충 음성인 110명의 혈청에서 1명이 네오포자충과 반응하였다. 교차반응을 보인 12 명의 혈청을 western blot으로 확인하였을 때, 톡소포자충 항원과는 다양한 양상으로 반응하였으나, 30 kDa (SAG1)과 22 kDa (SAG2) 항원과 강하게 반응하였다. 네오포자충과의 반응은 급격히 감소하였으나, 세 경우에서 43 kDa 단백질과 반응하였으며, 음성 혈청군의 1명도 43 kDa 단백질과 반응하였다. 면역형광법에서는 모든 양성 혈청이 톡소포자충의 세포막에 표지되었으나, 네오포자충과의 반응은 세포막, 세포 내 소기관, 혹은 둘 다를 표지하였다. 이로써, 톡소포자충과 네오포자충의 항원적 교차반응과 사람 혈청에서 네오포자충에 대한 항체의 존재를 확인하였으며, 네오포자충에 의한 인체감염 가능성에 대해서는 추후 연구가 필요할 것으로 판단된다. 또, N. caninum의 우리말 이름을 네오포자충으로 제안한다.

• 한국토양비료학회지
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• 제45권6호
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• pp.900-903
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• 2012

Cadmium is known to be very toxic to human health and can be relative easily translocated from soils in plants. Therefore, a rapid method for screening Cd in soils and crops has become more and more important. For this reason, we examined a rapid immunochromatograpy (ICG) test kit which uses antigen-antibody reaction based on immunoassay and chromatography. Soils and rice grains collected from mine waste-contaminated sites were determined for their Cd contents using this kit. For comparison purposes, 0.1 M HCl and ICP-OES were employed as a conventional extraction and determination method. Cadmium contents in rice grains determined using ICG technique were $0.46{\sim}2.39mg\;kg^{-1}$ and Cd contents determined using 0.1 M HCl and ICP-OES were $0.52{\sim}1.97mg\;kg^{-1}$. The correlation between these two Cd contents were statistically significant ($r^2$=0.930). The results of Cd contents in soils also showed a statistically significant relationship between these two methods ($r^2$=0.975). On the basis of these results, ICG technique can be applied to rapidly quantify Cd in crops and soils. However, further research is necessary to apply ICG technique for the field screening.

• Journal of Biosystems Engineering
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• 제32권6호
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• pp.440-447
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• 2007

In this study, a biosensor was developed using an enzyme-linked immunosorbent assay (ELISA) to rapidly measure the fungicide iprovalicarb residues in agricultural products. The biosensor was designed to include micro-pumps and solenoid valves for fluid transport, a spectrophotometer cuvet as a reaction chamber, a photodiode with a light-emitting diode for optical density measurement, and a control microcomputer to implement assay. The rate of change in optical density of the cuvet was read as final signal output. Micro-pumps were evaluated to investigate their delivery capability, the highest values of the error and the coefficient of variation were 4.3% and 4.6% respectively. As the incubation period was reduced from 15 minutes to 11 minutes to shorten the total processing time, the sensor sensitivity was decreased as the antibody dilution ratio was reduced to a half. The maximum usable period of the coated cuvet was found to be two days with 1% error limit. To predict the concentration of the iprovalicarb residue in agricultural products, a linear calibration model was obtained with r-square values of 0.992 for potato and 0.985 for onion. In validation test for the samples of potatoes and onions against the high performance liquid chromatography, very high correlation values were obtained as 0.996 and 0.993 respectively. Using the cuvet immobilized with antigen, it took 21-minutes for the biosensor to complete the measuring process of the iprovalicarb residues.

• 한국수의병리학회지
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• 제6권1호
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• pp.19-26
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• 2002

The purpose of this study was to investigate to potective effects against porcine epidemic diarrhea virus (PEDV) infection in piglets by administration of the PEDV antiserum orally at 2 hrs, 24hrs and 36hrs after birth. six piglets administered the antiserum were experimentally infected with PEDV at five-day-old. Control group were four piglets infected with PEDV only. Serum antibody titers against PEDV were examined by serum neutralization (SN) test, dectection for PEDV or PEDV antigen from feces and small intestines was tested by reverse transcription-polymerase chain reaction (RT-PCR) and indirect immunoflurescence (IFA). The results obtained were as follows; 1. The piglets administered the PEDV antiserum showed higher antibody titers than those of control group and sustained during the experimental period. 2. The detection rate of PEDV in feces and small intestines by RT-PCR were 26.2% and 16.7% in PEDV antiserum treated group and 48.1 % and 75.0% in control group, respectively. 3. The detection rate of PEDV antigen in the small intestine by IFA were 0% in PEDV antiserum treated group and 50.0% in control group, respectively. It was concluded that oral administration of antiserum against PEDV to piglets was effective in preventing PEDV infection.