• 한국동물위생학회지
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• 제19권1호
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• pp.30-38
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• 1996

Enzyme-linked immunosorbent assay (ELISA) was developed to detect rotavirus antigen in fecal samples using VP6-specific monoclonal antibody(2B12). The ELISA for rotavirus antigen detection found to have specificity to all bovine and porcine rotaviruses tested but not to bovine viral diarrhea virus and bovine coronavirus. The ELISA appeared to have similar sensitivity and specificity compared to fluorescence antibody assay(FA) and electropherotyping (PAGE).

• 대한수의학회지
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• 제31권2호
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• pp.223-228
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• 1991

Most progesterone enzyme immunoassays(EIA) are used liquid-phase double-antibody or single-antibody seperation. These methods consume considerable time and reagents because of the requirements for several washing and centrifugation steps involving the reactants. Because of these several problems, we were prompted to develop an effective enzyme-linked immunosorbent assay(ELISA) system that would be equal or superior to RIA for assay of progesterone. The results were obtained as follows. 1. Cross reaction of the progesterone antiserum with other steroids determined was shown with progesterone(100%), $11{\alpha}$-deoxycorti-costerone(2.271%), but the other steroids were shown below 0.9%. 2. Standard curve for progesterone ELISA was shown available difference according to progesterone concentration from 0 to 1,000pg/ml. 3. The lower limit of sensitivity was 0.2pg/well 4. Progesterone concentration was 1.6ng/ml for before parturition, and that was below 0.5ng/ml for after parturition. This development enzyme-linked immunosorbent assay for progesterone can be detected pregnancy diagnosis in cattle, and also applicable 10 research on physiological function including such as reproductive disorders.

• Journal of Microbiology and Biotechnology
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• 제14권3호
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• pp.612-619
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• 2004

A competitive direct enzyme-linked immunosorbent assay (cdELISA) was developed for selective and rapid detection of a glycopeptide antibiotic, teicoplanin (TP). TP was conjugated to bovine serum albumin (BSA) for use as an immunogen. Repeated subcutaneous injections of 0.5 mg of the conjugate was effective in generating specific polyclonal antibody (PAb) toward TP in rabbits, as determined by cdELISA. TP-horseradish peroxidase conjugate (TP-HRP) was used as an enzyme marker. The cdELISA was developed based on a competition reaction between TP-BSA PAb and TP-HRP conjugate. The TP-BSA PAb was highly sensitive (detection limit, 0.3 ng/ml and specific toward teicoplanin, showing no cross-reactivity to other glycopeptide antibiotics including vancomycin. There were good correlations ($r^2$=0.84 and 0.76, respectively) between cdELISA and microbiological assay, and high-performance liquid chromatography. The cdELISA system developed in this work is expected to be useful not only for selective and rapid monitoring of TP but also for study of TP pharmacokinetics.

• 약학회지
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• 제38권3호
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• pp.351-357
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• 1994

The levels of maternal immunity enhancing factor(MIEF), which is an immunomodulatory protein identified from bovine colostrum, were determined by indirect competitive enzyme-linked immunosorbent assay(ELISA) for the colostrum and normal milk collected during the first two weeks of lactation. The mean concentration of MIEF in the colostrum of the first day of lactation was $109\;{\mu}g/ml$, and fell from the third day of lactation to $3{\sim}4\;{\mu}g/ml$. The molecular weight of the purified MIEF determined by reducing SDS-PAGE and TSK G2000SW column chromatography was 22,000 and 24,000 daltons, respectively, showing that MIEF is a monomeric peptide in its native form. To examine the capacity of MIEF to induce differentiation of B Lymphocytes, human tonsillar Iymphocytes were cultured in the presence of different concentrations of MIEF, and then antibody secreting cells were enumerated by enzyme-linked immunospot(ELISPOT) assay. When added to cultures of human tonsillar Lymphocytes, MIEF induced differentiation of resting B Iymphocyte to antibody secreting plasma cells as efficiently as LPS.

• Archives of Pharmacal Research
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• 제23권6호
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• pp.548-553
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• 2000

Two drug-enzyme conjugates of dexamethasone-subtilisin and dexamethasone-cellulase have been synthesized and characterized to study their drug-protein incorporation ratio, immunoreactivity, enzyme activity and stability and these studies proved that a variety of drug enzyme conjugates could also be synthesized and characterized.

• 공업화학
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• 제25권5호
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• pp.544-547
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• 2014

본 연구에서는 효소면역 분석법(enzyme-linked immunosorbent assay)과 면역크로마토그래픽 기법을 결합하여 Legionella pneumophila 검출을 위한 면역스트립을 제작하였다. 면역스트립은 4종의 멤브레인을 이용하여 제작하였다. 니트로셀룰로오스 멤브레인은 포획항체를 고정화하여 신호 발생을 일으키기 위해 사용되었고, 두 종류의 유리섬유 멤브레인은 각각 중합체 패드와 시료주입 패드로 사용되었다. 셀룰로오스 멤브레인은 모세관 현상으로 시료흐름을 유도하는 흡수 패드로 이용하였다. 샌드위치 면역반응과 효소반응에 의해 30 min 이내에 생성된 발색신호는 정성 및 정량 분석이 가능하였다. 분석조건 하에서 육안에 의한 정성 검출뿐 아니라, $1.3{\times}10^3-1.3{\times}10^6CFU/mL$ 범위의 L. pneumophila 농도를 디지털카메라와 자체 제작된 소프트웨어를 이용하여 정량적으로 분석할 수 있었다.

• Korean Chemical Engineering Research
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• 제49권4호
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• pp.387-392
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• 2011

생촉매인 효소의 기질선택성은 다양한 분야에서 유용하게 이용되고 있다. 그 중에서도 효소결합면역흡착검사(enzyme-linked immunosorbent assay, ELISA)는 항원항체의 결합을 항체와 공유결합된 효소의 반응으로 나타냄으로써 다양한 항원들의 진단을 가능케 했다. 하지만 기존의 효소결합면역흡착검사는 하나의 항체당 하나의 효소가 결합된 형태이기 때문에 감도(sensitivity)의 증가 폭에 그 한계가 있으며, 이를 극복하기 위한 방안으로 하나의 항체당 결합된 효소의 수를 증가시킴으로써 혁신적인 감도의 향상을 가져오는 연구가 진행되었다. 최근 나노바이오촉매(nanobiocatalyst, NBC) 접근방식을 이용한 효소활성의 안정화는 효소결합면역흡착검사의 감도 향상뿐만 아니라 그 성능의 안정성을 확보할 수 있는 연구결과로 이어지고 있다. 본 총설에서는 일반적인 효소결합면역흡착검사의 기본적인 원리와 감도향상을 위한 연구, 그리고 성능안정성(performance stability)을 향상시키기 위한 나노바이오촉매-결합면역흡착검사(Nanobiocatalyst-Linked Immunosorbent Assay, NBC-LISA)에 대하여 살펴보고자 한다.

• 공업화학
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• 제22권5호
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• pp.522-525
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• 2011

본 연구에서는, enzyme-linked immunosorbent assay (ELISA)와 면역크로마토그래픽 기법을 결합하여 Staphylococcus aureus 검출을 위한 면역스트립을 제작하였다. 면역스트립은 4개의 서로 다른 기능을 가진 멤브레인을 이용하여 만들어졌다. 니트로셀룰로오스 멤브레인은 항체와의 결합력이 높기 때문에, 포획항체를 고정화하였고, 다공성 멤브레인들을 통하여 모세관 현상으로 인해 시료흐름을 유도하였다. 효소반응에 의해 생성된 발색신호는 디지털카메라와 자체 제작된 소프트웨어를 이용하여 정성, 정량분석 하였다. 최적의 분석조건 하에서 30 min 이내에 $2.7{\times}10^4{\sim}2.7{\times}10^7CFU/mL$ 범위의 S. aureus 농도를 측정할 수 있었다.

• Bulletin of the Korean Chemical Society
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• 제31권2호
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• pp.384-388
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• 2010

Human microsomal prostaglandin E synthase-1 (mPGES-1) catalyzes the conversion of prostaglandin $H_2$ ($PGH_2$) into prostaglandin $E_2$ ($PGE_2$). To establish a stable and efficient method to assess the activity of mPGES-1, a coupled enzyme assay system using mPGES-1, 15-hydroxyprostaglandin dehydrogenase (15-PGDH) and phosphomolybdic acid (PMA) was developed. In this assay system, $PGH_2$ was converted to $PGE_2$ by mPGES-1, and then $PGE_2$ was further transformed to the 15-keto-$PGE_2$ by 15-PGDH accompanying the production of NADH, which was easily detected by fluorescence spectrometry in a multi-well plate format. During the reaction, spontaneous oxidation of $PGH_2$ was prevented by PMA. Using this novel assay, the $K_m$ value of mPGES-1 for $PGH_2$ and the $IC_{50}$ value of the previously characterized inhibitor, MK-886, were determined to be 0.150 mM and $2.8\;{\mu}M$, respectively, which were consistent with the previously reported values. In addition, low backgrounds were observed in the multi-wall plate screening of chemical compounds.

• 한국미생물·생명공학회지
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• 제16권3호
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• pp.226-230
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• 1988

Trichoderma viride가 분비하는 cellobiohydrolase를 항원으로 사용하여 이 섬유소 분해효소에만 특이적인 항체를 얻을 수 있었다. 이 항체를 이용하여 column single immunodiffusion 방법에 의해서 cellobiohydrolase의 농도 2-10 $\mu\textrm{g}$ 범위 내에서는 직선적으로 정량 할 수 있었고, ELISA 방법을 사용할 때는 $10^6$의 항체 희석시는 40-110ng, $10^5$ 항체 희석시는 100-1,200pg 농도범위의 cellobiohydrolase 를 정량 할 수 있었다.