• Title/Summary/Keyword: ciELISA

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An Enzyme-Linked Immunosorbent Assay for Detection of Pantothenic Acid (판토텐산의 분석을 위한 효소면역측정법)

  • Shon, Dong-Hwa;Park, Youn-Sick;Bae, Gun-Won
    • Korean Journal of Food Science and Technology
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    • v.32 no.5
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    • pp.1009-1014
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    • 2000
  • In order to detect pantothenic acid (PA), conditions for enzyme-linked immunosorbent assay (ELISA) were established. Anti-PA-BSA antibody was produced from rabbits immunized with PA-bovine serum albumin (BSA) conjugates which were prepared by the bromoacetyl chloride [Bc] method (PA-BSA[Bc]) and by the periodate oxidation [Po] method (PA-BSA[Po]). PA-BSA[Bc] and PA-BSA[Po] was used as a coating antigen for competitive indirect(ci)ELISA. The Anti-PA-BSA[Po] antibody on ciELISA showed no competitive reaction. The detection limit of PA by ciELISA using Anti-PA-BSA[Bc] antibody was 1 ppm. The Anti-PA-BSA[Bc] antibody showed little cross-reactivity to PA derivatives such as pantoyllactone, pantetheine, pantothenyl alcohol, and acetyl CoA. The detection limit of PA by microbiological assay (MBA) was 10 ppb. Assay recoveries of PA in egg, cow's liver, and lettuce by ciELISA were 109, 64, and 344%, respectively, comparing with the MBA results.

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Immunological Assay to Detect Irradiated Beef

  • Lee, Ju-Woon;Yook, Hong-Sun;Lee, Hyun-Ja;Kim, Jung-Ok;Byun, Myung-Woo
    • Preventive Nutrition and Food Science
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    • v.6 no.2
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    • pp.91-95
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    • 2001
  • Competitive indirect enzyme linked immunosorbent assay (Ci-ELISA) was used to obtain the preliminary data for the detection of irradiated beef. Ci-ELISA was individually formatted with polyclonal antibodies produced from 2 kinds of bovine proteins, myosin and bovine serum albumin (BSA). Beef round, loin and tender loin were vacuum-packaged and subdivided into 3 groups of 1) irradiation; 2) irradiation and chilled at 4$^{\circ}C$ for 7 day; 3) irradiation and frozen at 2$0^{\circ}C$ for 2 months to observe the changes under different storage and/or distribution conditions. Irradiation was performed at 3, 5 and 7 kGy. Protein solutions prepared from the sample were tested by formatted Ci-ELISA. Detected concentrations of myosin and BSA decreased with the increased irradiation dose in all samples with different reduction rates. Myosin was more susceptible to freezing than BSA. Samples irradiated at 5 kGy or above could be differentiated from non-irradiated ones by Ci-ELISA. These results indicate that immunological assay can be used as a detection method for irradiated beef.

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Development of Competitive Indirect ELISA for the Detection of Buckwheat in Processed Foods (가공식품 중 메밀 검출을 위한 경합 ELISA의 개발)

  • Back, Su-Yeon;Do, Jeong-Ryong;Shon, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.46 no.3
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    • pp.269-275
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    • 2014
  • We developed a competitive indirect enzyme-linked immunosorbent assay (ciELISA) for determining the buckwheat content in processed foods by using rabbit polyclonal antibodies against buckwheat proteins (BWP). The detection limit of this assay was $0.05-100{\mu}g/mL$. The cross-reactivities of the anti-BWP antibodies toward BWP, buckwheat flour, whole buckwheat, and cereals (wheat flour, whole wheat, black bean, mung bean, red bean, brack rice, brown rice, glutinous rice, white rice, millet, African millet, nonglutinous millet, adlay, and rye) were 100, 17.9, 11.8, and 0%, respectively. Thus, the antibodies were found to be specific for buckwheat only. When buckwheat flour was heated for 30 min, the mean assay recoveries of BWP were 83.0% at $60-90^{\circ}C$ and 44.5% at $100^{\circ}C$. The spike test showed that the mean assay recoveries of buckwheat from raw noodle, boiled noodle, starch gel, and cereal flour were 99.1, 98.6, 81.1, and 104%, respectively. For the 22 commercial items tested, the qualitative coincidence ratio of assay result and the corresponding value indicated on the item's package label was 100%. However, the average quantitative coincidence ratios from 12 commercial items were 31.6%. Thus, the results suggest that ciELISA is an efficient tool to detect buckwheat in processed foods.

Comparison of Two Commercial Antibody Enzyme-Linked Immunosorbent Assays for Detection of Porcine Reproductive Respiratory Syndrome Virus Infection (돼지생식기호흡기증후군(PRRS) 바이러스 감염 항체 검출 ELISA 상용 키트의 정확도 비교)

  • Pak, Son-Il;Lee, Seung-Hwan;Park, Kyung-Ae
    • Journal of Veterinary Clinics
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    • v.33 no.2
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    • pp.102-106
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    • 2016
  • More than 20 years after the first report of porcine reproductive and respiratory syndrome virus (PRRSV) in Korea, the disease is still having major impact on domestic pig health and relevant industries. Although ELISA tests are commonly used by veterinarians to guide herd management, data on diagnostic performance of the test in field settings are very limited. The objective of this study was to evaluate two commercially available PRRSV ELISA (IDEXX PRRS X3 ELISA and Bionote PRRSV ELISA 4.0) to detect antibodies against PRRSV on serum samples. To this end, a total of 1,108 sera were recruited from 35 swine farms located in Gyeonggi province and tested at the Gyeonggi Province Veterinary Service Center. All tests were performed according to the manufacturer's instructions, by laboratory technicians who routinely perform PRRS testing on blood samples. Samples were collected from two sources of swine populations with different PRRS prevalence; 60 samples (5.4%) were originated from breeding farms and the remaining 1,048 samples (94.6%) were from farrow-to-finish farms. We applied Bayesian latent class model (LCM) for two-tests in the two-population when the accuracy of the gold standard is not available. The model estimated that Bionote ELISA was a bit more specific but slightly less sensitive. The estimated sensitivity and specificity of the IDEXX ELISA were 99.8% (95% CI 98.1-100%) and 86.4% (95% CI 81.4-96.5%), respectively. Sensitivity, specificity, positive predictive value and negative predictive value for Bionote kit were 98.7% (95% CI 92.8-100%), 89.8% (95% CI 86.2-93.1%), 93.8% (95% CI 91.5-96.0%), and 97.8% (95% CI 87.1-100%), respectively. Based on the Bayesian 95% credible intervals, the sensitivity and specificity of the two ELISAs were not significantly different each other when assuming that two kits were imperfect, indicating that two kits performed equally well in terms of sensitivity and specificity in our filed setting.

Enzyme-linked Immunosorbent Assay for the Detection of Hen's Egg Proteins in Processed Foods

  • Shon, Dong-Hwa;Kim, Hyun-Jung;Kim, Soo-Ho;Kwak, Bo-Yeon
    • Food Science of Animal Resources
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    • v.30 no.1
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    • pp.36-41
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    • 2010
  • The Hen's egg is widely used in many processed foods as an ingredient and is one of the most prevalent food allergens in children. To detect egg proteins in processed foods, we developed a competitive indirect enzyme-linked immunosorbent assay (ciELISA) using an anti-ovomucoid (OM) antibody, which was produced by immunization of rabbits with OM, the most heat-stable component of the egg proteins. The detection limit of this quantitative assay system was 30 ng/mL. Cross-reactivity of the anti-OM antibody toward OM, ovalbumin, skim milk, casein, whey protein isolate, and isolated soy protein was 100, 0.4, 0.2, 0.04, 0, and 0%, respectively. In the spike test of egg white powder in milk replacer, commercial sausage, and in-house sausage, the assay recoveries ($mean{\pm}SD$) were $129{\pm}13.7%$, $73.9{\pm}12.5%$, and $65.5{\pm}13.6%$, respectively. When egg white in a commercial crab meat analog and sausage was determined by ciELISA, the assay recovery was found to be 108% and 127%, respectively. The combined results of this study indicate that this novel ciELISA for OM detection could be applied for the quantification of hen's egg proteins in processed foods.

Lipopolysaccharide Yields from Rhodobacter capasulatus with indirect ELISA

  • Yoo, Tae-Eun;Lee, Hyun-Soon
    • Journal of Microbiology
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    • v.34 no.3
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    • pp.255-262
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    • 1996
  • The lipopolysaccharide (LPS) yields were measured in Rhodobacter capsulatus under several conditions by the ELISA method. The purification of LPS was done by affinity chromatography of IgG coupled CNBr-activated sepharose-4B instead of ultra-centrifugation. The purity of the LPS didn't show much difference between affinity chromatography and ultra-centrifugation method, but affinity chromatography method required much fewer organisms and was more convenient. LPS yield was measured in ng units by the ELISA method. Mannitol was a better single carbon source than other sugars, but mixing two carbon sources resulted in greater LPS yields than any sugar alone. LPS yield was directly proportional to $NH_ 4CI$ concentration, with optimum yields at 0.05% nitrogen. In contrest to LPS yields, which decreased at 0.005% nitrogen concentration total protein was increased 16 times. Calcium influenced LPS yields. At 0.7 mM $CaCI_ 2$, the LPS yield was 16.5 $\mu$g/mg DW, five times the yield without calcium.

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Property Comparison of Polyclonal Anti-Soy Protein Antibodies Produced for ELISA (효소면역측정법을 위한 다클론 항대두단백 항체의 생산 및 특성비교)

  • Shon, Dong-Hwa;Kim, Hyun-Jung;Yun, Sung-Seob
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1221-1226
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    • 2000
  • Specific antibodies were produced to develope the enzyme-linked immunosorbent assay for analysis of soy proteins and the properties of the antibodies were compared. Isolate soy protein(ISP), and ISP heated with SDS and urea (ISP(SU)), acidic subunits(AS) of 11S globulin were immunized to produce polyclonal antibodies. By using competitive indirect ELISA(ciELISA), the reactivities of the antibodies toward soy proteins treated with different methods were investigated and shown as $IC_{50}$. $IC_{50}'s$ of anti-ISP antibodies to ISP, ISP(SU), ISP treated with 2-ME(ISP(ME)), and crude 11S were 20, 30, 36, and $1000\;{\mu}g/mL$, respectively. And the values of anti-ISP(SU) antibodies to the same antigens were 100, 5, 4, and $220\;{\mu}g/mL$ and those of anti-AS antibodies were 20, 2, 2.5, and $200\;{\mu}g/mL$, respectively. Therefore, anti-AS antibodies showed the highest reactivities toward soy proteins among the produced antibodies as determined by ciELISA.

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