• Korean Journal of Animal Reproduction
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• v.15 no.2
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• pp.149-155
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• 1991

본 실험은 H-Y 항원 유전자 크로닝을 위한 기초연구로서 H-Y 항원의 특성을 규명하기 위하여 친화성 크로마토그래피에 의하여 H-Y 항원을 분리·정제하였다. 정소 추출액을 항체가 결합된 column에 결합시킨 뒤 10% acetic acid로 용출시켰다. 용출된 분획을 모아 농축한 후 HPLC와 SDS-PAGE를 실시하여 H-Y 항원의 분자량은 약 6,7000달톤 임을 알 수 있었으며 isoelectric focusing에 의하여 등전점(pI)은 5.0인 것으로 측정되었다. H-Y 항원에 대한 단일클론항체와 표지항원으로는 H-Y 항원-ABEI(aminobutylethyl isoluminol)를 사용하여 H-Y 항원 정량을 위한 화학발광면역분석법을 개발하였다. 항원항체 반응후 빛의 측정은 NaOH 존재하에서 microperoxidase/H2O2를 이용한 산화반응으로 실시하여 10초간 측정한 빛의 양을 적분하였다. H-Y 항원의 농도와 빛의 양과는 역비례하였으며 감도는 11.8ng/tube 정도이었다.

• Journal of Dairy Science and Biotechnology
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• v.15 no.1
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• pp.33-44
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• 1997

We investigated the immunological function of cheese whey protein concentrate (CWPC), which is a by-product of cheese production, using mitogenic activity in murine splenocytes as an index. A fraction isolated by gel filtration and anion exchange chromatography of CWPC showed high mitogenic activity, comparable to the activity of lipopolysaccharide (LPS). The fraction was detected as a single band on SDS-PAGE. It contained calcium, inorganic phosphorus, and carbo-hydrate, indicating the active component to be a glycophosphopeptide (GPP) Since pronase digestion of GPP did not reduce its mitogenic activity, carbohydrate rather than peptide may be important in the activity, When applied on an anti-${\beta}$-caseinophosphopeptide (${\beta}$-CPP ) antibody affinity column, the GPP was separated into two components, one with affinity to ${\beta}$-CPP and the other without such affinity. Both the components contained N-linked oligosaccharide chains and had the mitogenic activity. These results demonstrate that cheese whey contains a GPP having strong mitogenic activity

• Biomedical Science Letters
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• v.2 no.1
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• pp.41-48
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• 1996

The C-reactive protein(CRP) from ascitic and pleural fluid was purified using calcium dependent affinity chromatography of CNBr activated Sepharose-4B covalently coupled to p-diazonium phenylphosphorylcholine(DPPC) and hydroxylapitite chromatography. Polyclonal antibody was prepared from rabbit by immunizing the purified CRP. Specific immunoglobulin G was isolated using affinity chromatography and coupled to microparticles. A sensitive microparticle-based immunoassay was developed to measure CRP within 3 mins. The detection range was between 0.5mg/dl) and 20mg/dl in serum, showing strong response in the range of 0.7~2.9 mg/dl, weak response in 5.0~13.2 mg/dl and zone phenomenon over 28mg/dl. The average value of CRP in 74 samples was 3.8mg/dl and most of the values were lower than 10mg/dl .The CRP values of serum samples were determined by our microparticle-based immunoassay, and were compared with those obtained using the other commercial products(B Co., France and I Co., Japan). Good correlations were shown between the values obtained by our developed microparticle-based immunoassay system and those by other commercial products. All performance characteristics evaluated make our developed microparticles-based immunoassay suitable for a simple, rapid, and reliable screening of CRP in serum.

• Applied Chemistry for Engineering
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• v.25 no.5
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• pp.544-547
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• 2014

In this study, an enzyme-linked immunosorbent assay (ELISA) and immuno-chromatographic technique were combined to fabricate immuno-strip sensors for the detection of Legionella pneumophila. The immuno-strip sensor was manufactured with four different membranes. A nitrocellulose membrane was used to immobilize capture antibody and generate signals due to the high affinity to antibodies, and glass fiber membranes were used as a conjugate release pad and a sample application pad. A cellulose membrane was used as an absorption pad to induce sample flow by the capillarity. Colorimetric signals produced by sandwich immuno-reaction and enzyme reaction could be analyzed qualitatively and quantitatively within 30 min. Under the given experimental conditions, sensor signals with L. pneumophila samples were observed qualitatively by naked eyes and measured quantitatively in a range of $1.3{\times}10^3-1.3{\times}10^6CFU/mL$ with a digital camera and home-made image analysis software.

• Proceedings of the Korean Society of Food Hygiene and Safety Conference
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• 2002.05a
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• pp.188-191
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• 2002

• Journal of Food Hygiene and Safety
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• v.37 no.1
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• pp.15-20
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• 2022

The purpose of this study was to compare the conventional culture method, enzyme immune method and the PCR method using species-specific primer in the analysis on the Salmonella spp. found in domestically distributed pet foods. For the study, Salmonella spp. were detected from 175 samples. From the conventional culture method and the PCR method, two samples (jerky and corn gluten) were determined as positive. Also, from the enzyme immune method, one sample (corn gluten) was test-positive. The study revealed that application of the PCR method with species-specific primer allows better distinguishment between the species of the strain collected from the samples than the conventional culture method and/or the enzyme immune method.

• Proceedings of the Korean Society of Food Hygiene and Safety Conference
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• 2004.05a
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• pp.252-252
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• 2004

• Microbiology and Biotechnology Letters
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• v.20 no.2
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• pp.225-232
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• 1992

In order to develop an enzyme-linked immunosorbent assay(ELISA) for detecting aflatoxin $B_1(AFB_1)$, we produced and purified antibodies, thereafter established and evaluated methods of direct and indirect competitive ELISA. Anti-AFB, antisera, produced by immunizing rabbits with $AFB_1$-1-(0-carboxymethy1)oxime-bovine serum albumin conjugate ($AFB_1$-BSA), were removed of anti-BSA antibodies by quantitative precipitation reaction and further purified by ammonium sulfate precipitation and DEAE-Sephadex A-50 ion exchange chromatography. Purified IgG fractions were used as anti-$AFB_1$ antibodies. The antibodies, whose titer was deterrnined extremely high above $2 \times 10^6$, showed low cross-reactivity of 3~34% against $AFB_1$ analogues such as G2, B2, and GI. From the standard curves of direct and indirect competitive ELISA for AFBI, the detection ranges were found 0.2~20 and 1~10, 000 ng/ml(ppb) respectively. In their sensitivity, stability, simplicity, and rapidity, the direct method was more suitable than the indirect method for practical use.

• Korean Journal of Animal Reproduction
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• v.11 no.1
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• pp.42-62
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• 1987

본 실험은 성숙가토의 정자와 정장의 항원성을 조사하여 면역적 불임원인 규명에 필요한 기초자료를 얻고자 시도하였다. 인공질로 채취한 정액을 원심분리하여 얻은 정자와 정장을 항원으로 사용하였으며 항원성의 측정방법은 크로마토그라프에 의한 단백질 분리, SDS-PAGE, HPLC, 한천확산법, 전기영동, 수동적혈구응집방법 및 부동화시험이었다. 얻어진 결과는 다음과 같다. 1. SDS-PAGE 전기영동시 정상가토 정장에서 약 23개의 단백질이 분리되었고, 그중 분자량이 약 20,000되는 단백질부분이 정관절제 수술한 정장에서 나타나지 않았다. 2. 정상가토 정장을 HPLC를 이용한 분석에서 3개의 peaks을 볼 수 있었고, 정관절제수술한 것에서는 peak 1에 해당하는 단백질이 소실되었다. 3. 한천확산시험에서 정상정장은 이종혈청과는 4개의 침강선을 나타냈고, 전기영동에서는 7개의 침강선을 나타냈다. 4. 이종면역에서 정자 및 정장은 항체가 상승이 용이하였지만, 동종면역시는 추가면역이 필요하였으며 개체간의 역가차이를 보였다. 정자면역한 자성가토에서는 수동적혈구응집반응을 나타냈지만, 한천확산 및 전기영동반응은 보이지 않았고, 같은 처치를 받은 웅성가토에서도 역시 같은 반응양상을 나타냈다. 5. 동종 및 이종항혈청을 이용한 교차전기영동방법으로 정장의 항원적 구성요소를 구별할 수 있었다. 사출된 정액에서 분리된 정장은 일부 항원을 포함하고 있었으며 이는 성숙한 정자에서 기인한 것으로 사료되었다.

• KSBB Journal
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• v.15 no.3
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• pp.243-246
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• 2000

One of recent topics in the case of hepatits B virus(HVB) is the value of hepatitis B surface antigen(HBsAg) concentration as a prognostic maker. We developed uantitative method of rapid immunochromatographic assay(ICA) kit for HBsAg using computer image analysis (CIA) for the purpose of home diagnosis. uantitative ICA using CIA demonstrated integrated optical density(IOD) values proportional to log of reference HBsAg concentrations in the range of 2-200 ng/mL and enzyme-linked immunosorbent assay(ELISA) demonstrated the same in the range of 0.1-100 ng/mL however the test results with sample sear showed the same concentration on both kits. Furthermore repeated tests with the same samples revealed that this quantitative ICA using CIA would be reproducible and coefficient of variation(CV) of the results was 1.38~6.30%.