• Title/Summary/Keyword: 면역분석법

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Production and Characterization of the Monoclonal Antibody for Human Serum Albumin (사람 혈청 알부민에 대한 단서포군 항체의 제조와 특성 연구)

  • 태건식;안혜진
    • Proceedings of the KAIS Fall Conference
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    • 2000.10a
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    • pp.243-244
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    • 2000
  • Serum albumin은 혈청 단백질 중 50-60%를 차지한다. 알부민은 순환계 내에서 삼투압을 유지시켜 세포내외의 체액을 유지 및 지용성 물질치 이동에 관여한다. 신사구체와 기저막의 투과성이 증가하거나 세뇨관의 흡수감소 등으로 단백질의 요배설량이 하루에 150mmg 이상되는 단백뇨는 신장내의 병변이 있음을 나타내는 중요한 지표이다. 이러한 단백뇨는 크게 사구체성 단백뇨, 세뇨관성 단백뇨, 혼합성 단백뇨로 나눌수 있는데 이중 사구체 기저막의 손상으로 발생되는 사구체성 단백뇨는 알부민과 같은 고분자량의 단백질이 소변 내로 배설되는 현상이다. 일반적으로 알부민이 정상치 이상으로 소변을 통하여 배설되는 것을 'microalbuminuria'라고 하며 당뇨병으로 인한 신장병변을 예견하는데 중요한 지표로 알려져 있다. Microalbuminuria는 방사 면역확산법, 방사성 면역측정법, 면역혼탁측정법, 비탁측정법, 효소결합면역측정법(ELISA) 등으로 검출할 수 있으나 최근에는 안전하면서도 민감도가 높고 측정이 용이한 방법인 ELISA를 이용한 면역분석방법이 많이 사용되고 있다. 면역특이성이 떨어지는 다세포군 항체를 사용한 방법의 문제점을 극복하기 위해서 세포융합기술을 이용한 단세포군항체를 생산하는 세포주를 개발하였고 그 특성을 규명하였다. 따라서 본 연구에서 세포융합기술을 도입하여 개발된 인간 혈청 알부민에 대한 단세포군항체는 glycohemoglobin에 대한 단세포군항체와 더불어 당뇨병의 진행 정도를 진단할 수 있는 키트의 원료로 사용될 수 있다.

Bioluminescence immunoassay for neurotransmitter, serotonin using aequorin as a Label (Aequorin을 표지물질로 사용한 신경전달물질, 세로토닌에 대한 생물발광면역분석법)

  • Ryu, Ji-Eun;Choi, Hee-Seon;Park, Ho-Young;Rhee Paeng, In-Sook
    • Analytical Science and Technology
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    • v.23 no.1
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    • pp.60-67
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    • 2010
  • A sensitive competitive heterogeneous bioluminescence immunoassay for serotonin was developed using photoprotein, aequorin as a label for the first time with the optimal assay conditions; especially, serotoninavidin conjugate was prepared by Mannich reaction and the synthetic process of serotonin-avidin conjugate was optimized by controlling the initial molar ratios of serotonin, formaldehyde and avidin (1:12,000:25). The developed bioluminescence immunoassay for serotonin showed good sensitivity (LOD of 0.68 ng/mL) with wide area of dynamic range ($5.0{\times}10^{-10}\;M\sim5.0{\times}10^{-7}\;M$). (cf. the range for serotonin in human blood serum is $151{\pm}45\;ng$/mL). In addition, cross-reactivity studies demonstrated that 5-methoxytryptamine showed some cross-reactivity (28.0%), whereas 3-methylindole, melatonin and 5-hydroxylindole-3-acetic acid showed no crossreactivity, and good recoveries were obtained in serum. Thus, this developed method provides a good tool to monitor serotonin in serum.

A feasibility study on the Application of Europium in the Lateral Flow Assay System with Time Resolved fluorometry (유러피엄을 이용한 측방유동 시분할 형광 분석기법)

  • Jung, JIn-Ha;Nam, Ki-Bong;Ko, Dong-Sup;Moon, Jeong-Dae;Jung, Dong-Suk;Choi, Yeoi-Ryul;Kim, Jae-Hoon
    • Proceedings of the Optical Society of Korea Conference
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    • 2004.07a
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    • pp.296-297
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    • 2004
  • 시분할 형광 분석법은 발광하는 형광을 통해 시료내의 미량의 물질을 분석 할 수 있는 분석법으로 생화학, 면역학 등 여러 분야에서 응용되고 있으며 새로운 응용 분야에 대한 연구가 활발하게 진행되고 있다. 본 연구에서는 유러피엄(Europium) 화합물을 면역학적 검출기로 사용한 측방유동 시료(lateral flow assay)를 시분할 형광 분석법으로 측정한 결과를 소개한다.

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Enzyme-Linked Immuno-Sorbent Assay for Bovine Caseins (우 Casein의 면역효소분석법)

  • 염행철
    • Korean Journal of Animal Reproduction
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    • v.16 no.2
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    • pp.87-102
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    • 1992
  • A rapid, sensitive, and specific enzyme-linked immuno-sorbent assay (ELISA) for bovine casein was developed. Biotinylated casein and peroxidase-conjugated avidin were used in the assay with antibody separated from yolks of immunized hens. Caseins were biotinylated with sulfo-N-hydroxy succinimido biotin and peroxi-dase-conjugated avidin bound the biotinylated casein which became bound to immobilized anti-body on a microplate. The antibodies were specific for bovine $\alpha$- and $\beta$-caseins, and their cross-reactivities with whey proteins, IgG, and serum albumin from bovine were not detectable by ELISA and Western blot. Various sensitivities ranging from 2ng/ml to 20${\mu}\textrm{g}$/ml of casein were achieved, and were controlled by adding vanous concentrations of the biotinylated casein. Parallelism was observed between standard and sample curves. The coefficients of variation of intra-assays and inter-assays from the most sensitive assay were 5.5 and 5.7%, respectively, at the 50% displacement. Casein contents of peripaturient milk samples showed that casein secretion rapidly increased 3d prepartum.

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Recent Progress in Multiplexed Detection of Biomarkers Based on Quantum Dots (양자점 기반 다중 바이오마커 검출법의 연구동향)

  • Kim, Yerin;Choi, Yu Rim;Kim, Bong-Geun;Na, Hyon Bin
    • Applied Chemistry for Engineering
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    • v.33 no.5
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    • pp.451-458
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    • 2022
  • Semiconductor quantum dots (QDs) are optical probes with excellent fluorescence properties. Therefore, they have been applied to various bio-medical imaging techniques and biosensors. Due to the unique optical characteristics of wide absorption and narrow fluorescence energy bands, multiple types of signals can be generated by the combination of fluorescence wavelengths from different QDs, which enables the simultaneous detection of more than two biomarkers. In this review, the advantages and applications of QDs and QD nanobeads (QBs) in multiple biomarker assays were described, and new developments or improvements in multiplexed biomarker detection techniques were summarized. In particular, recent reports were summarized, focusing on the design strategies in immunoassay construction and signal transducing materials for fluorescence-linked immunosorbent assays using QDs and immunochromatographic assays using QBs. New detection platforms will be developed for early diagnosis of diseases and other fields if multiplexed detection technologies of excellent accuracy and sensitivity are combined with artificial intelligence algorithms.

Production of Monoclonal Antibodies to Phytohormones and Its Application (식물홀몬에 대한 단클론성 항체 생산과 이용)

  • 황태익
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.34 no.s01
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    • pp.48-54
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    • 1989
  • An immunoassay techniques using monoclonal antibodies have been developed of the quantitative analysis of endogenous hormones in plants. In order to be useful for immunoassay, both a high degree of specificity and a high affinity are required. A system is described for production of hybridomas which secrete antibodies against the plant hormone. Using the system we were able to produce hybridmas with the desired antibody specificity by cell fusion and culture method. For a number of obvious reasons, monoclonal antibodies(mAb) were superior to polyclonal antibodies.

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Establishment of I-131, Tc-99m Labeling Methods to In-house Anti-CEA Antibodies and Evaluation of the Immunological Characteristics (국산 항 CEA 항체의 I-131, Tc-99m 표지법 확립 및 면역학적 특성 분석)

  • Chung, June-Key;Lee, Dong-Soo;Lee, Myung-Chul;Chung, Hong-Keun;Koh, Chang-Soon;Hong, Mee-Kyoung;Choi, Seok-Rye;Seo, Il-Taek;Chung, Jun-Ho
    • The Korean Journal of Nuclear Medicine
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    • v.26 no.2
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    • pp.346-354
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    • 1992
  • Cancer cells have several tumor-associated antigens on the cell surfaces, and antibodies against these antigens have been developed by many investigators. Radiolabeled antibodies have been used as new methods to diagnose and treat malignant tumors. Especially anti-carcinoembryonic antigen (CEA) is the most popular antibody for these purposes. In this investigation, we tried to label $^{131}I$ and $^{99m}Tc $ to anti-CEA monoclonal antibodies which were developed in the Seoul National University College of Medicine. We found CEA-79 and CEA-92 antibodies had the better immunological characteristics among 8 anti-CEA monoclonal antibodies. And radioiodination of CEA-79 could be performed by chloramine-T method, while radioiodination of CEA-92 by iodogen method. To label these antibodies with $^{99m}Tc $, we used pretargeting transchelation as direct labeling method. At first, $^{99m}Tc $ was bound to glucaric acid, and monoclonal antibody was reduced by $\beta-mercaptoethanol$. When these were incubated together. $^{99m}Tc $ bound to glucarate was switched to monoclonal antibody because of higher affinity. We established conditions of several steps in this method. Anti-CEA monoclonal antibodies labeled with $^{131}I$ and $^{99m}Tc $ are expected to be used valuably in the detection and treatment of malignant tumors.

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Studies on the improvement of reproductive efficiency in Korean native cows -Development of radioimmunoassay for progesterone- (한우(韓牛)의 번식효율(繁殖效率) 증진(增進)에 관한 연구(硏究) -Progesterone측정(測定)을 위한 방사선면역분석법(放射線免疫分析法)의 개발(開發)-)

  • Choi, Han-sun;Kang, Byong-kyu;Lee, Chung-gill;Son, Chang-ho
    • Korean Journal of Veterinary Research
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    • v.30 no.2
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    • pp.171-175
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    • 1990
  • Radioimmunoassay for progesterone in Korean native cows was developed using the anti-4-pregnen-$11{\alpha}$-ol-3,20-dione-hemisuccinate bovine serum albumin as an antibody. The final dilution of antibody was 1 : 5,000. The percentage recoveries of [$^3H$] progesterone was $83.5{\pm}1.6$. The sensitivity of the assay was 8 pg per ml. The intra- and interassay coefficients of variation for the quality controls were 6.2 and 10.4%, respectively. This radioimmunoassay can be applied to puberty, estrus dectection, pregnancy diagnosis, postpartum period and diagnosis of reproductive disorders in domestic animals.

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