• Korean Journal of Food Science and Technology
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• v.32 no.5
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• pp.991-996
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• 2000

Enzyme-linked immunosorbent assay was developed for the analysis of soy protein in foods. Competitive indirect ELISA (ciELISA) was established by using specific antibodies against the heat-stable acidic subunits (AS) of glycinin. Soy proteins in each sample used in this study were solublized in the presence of urea and DTT and boiled at $100^{\circ}C$ for 1hr and then were renatured with a cystine-containing solution. After these treatments, each isolated soy protein (ISP) heated at 60, 70, 80, $90^{\circ}C$ for 10 minutes showed almost the same curve as unheated one in the ciELISA. The detection limit of ISP was 0.3 ${\mu}g/mL$. Anti-AS antibodies have very low reactivities less than 0.1% toward non-meat proteins such as skim milk and casein and did not show any reactivities toward egg white powder and ovalbumin. When laboratory-made sausages containing ISP of $0.5{\sim}3%$ were assayed by ciELISA, the mean recovery was about 83% (C.V., 19%). In addition, the average content of soy protein in commercial sausages was 1.27%.

• YAKHAK HOEJI
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• v.45 no.2
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• pp.180-189
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• 2001

We have reported that water-extracted Korean mistletoe (KM-110) had various biological activities such as antitumor and immunomodulatory activity, and the pectin fraction (KML-C) of the extract was one of major factors related to its biological functions. In this paper, we produced murine monoclonal antibody (mAb) against KML-C. The cAbs obtained were largely classified into two groups according to specificity to KML-C and ML-I, a pectin from European mistletoe. One group mAbs (9H7-D10 and 3C2-lH4) strongly reacted with KML-C, but not ML-I. In contrast, another group cAbs (8Bll-2C5, BE12-3E9 and 5E10-Fl) reacted with both KML-C and ML-1. The subisotypes of these mobs were shown to be IgGl (9H7-lD10, 3C2-lH4 and 8Bll-2C5) or IgM (8E12-3E9 and 5E10-Fl). To develop an assay system for determination of the amount of KML-C, we established the sandwich ELISA (enzyme-linked immunosorbent assay) method using these mAbs and horse radish peroxidase (HRP)-labelled cAbs. In various combinations of the cAbs for coated antibody and detection antibody, the sandwich ELISA quantitatively detected KML-C, showing the detection limit ranging from 7-5,000 ng/ml. Especially reproducibility (C.V) of the sandwich ELISA, in which 8E12-3E9 was used for coating antibody and 8Bll-2C5-HRP for detection antibody, was 4.59-5.83 in intra assay, and 3.9-9.4 in inter assay.

• Korean Journal of Food Science and Technology
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• v.46 no.5
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• pp.538-543
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• 2014

A sandwich enzyme-linked immunosorbent assay method (sELISA) for detecting the presence of shrimp in processed foods was developed using rabbit polyclonal antibodies against tropomyosin produced by black tiger prawns (shrimp). Based on the standard curve derived using this method, the detection range of shrimp was determined to be $1-100{\mu}g/mL$. The cross-reactivity of these antibodies toward black tiger prawns, fleshy prawns, cocktail prawns, lobster, and blue crab was 100, 73, 155, 18, and 0%, respectively. When shrimp was heated for 10 min, the mean assay recovery of tropomyosin was 121-221% at $70-100^{\circ}C$ and 7.8% at $121^{\circ}C$. When shrimp was added to cream soup, weaning food, sausage, fish paste, and sauce, the mean assay recovery was 397, 639, 168, 234, and 0%, respectively. In sample tests involving 14 commercial items, the coincidence ratio of assay results and reference was 79%.

• Korean Journal of Veterinary Research
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• v.37 no.3
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• pp.583-593
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• 1997

Indirect fluorescent antibody test(IFAT) and enzyme-linked imuunosorbent assay (IgG-ELISA) as serological diagnostic tools were conducted to evaluate the usefulness for diagnosis of canine babesiosis infected with Babesia gibsoni in domestic various dog breeds, american pit bullterrier, military shepherd, and mongrel dogs. The results obtained from this study were abstracted as follows. The nonionic detergent Triton X-100 and absorbent bio-bead $SM_2$ were useful reagents for the preparation of pure merozoite antigen of B gibsoni to be used in ELISA. The optimum reaction in ELISA was shown when the protein concentration of ELISA antigen was measured as 625ng/ml and the conjugate concentration was diluted into 1/6000 fold. The average OD value of ELISA in sera determined with negative responses in IFAT was measured as $0.255{\pm}0.051$(490nm) and the cut - off value of OD was determined as 0.399(490nm). The serum antibodies in both of IFAT and ELISA were detected on one week after artificially infected with B gibsoni and these high antibody titers, 512X in IFAT and 1024X in ELISA, were long lasted until 15 weeks after infection. The reproducibility of reaction and stability of the antigen absorbed microtitration polystyrene plate preserved in $4^{\circ}C$ refrigerator and $-20^{\circ}C$ freezer, respectively could be lasted until 135 days after storage. The positive rates in IFAT by dog breeds were shown 8.1%(60/744 heads) in mongrel dogs, 81.3%(78/96 heads) in american pit bullterrier and 15.6%(15/96 heads) in military shepherd, while the positive rate in ELISA shown 17.6%(131/744 heads) in mongrel dogs, 83.3%(80/96 heads) in american pit bullterrier and 36.5%(35/96 heads) in military shepherd, respiectively. In the total of 936 heads surveyed with IFAT and ELISA the positive rates in IFAT and ELISA were 16.4%(153/936 heads) and 26.3%(246/936 heads), respectivily. Agreement of reactions between IFAT and ELISA was shown 82.4% in 936 dog sera. The specificity and sensitivity of ELISA reaction were 83.5% and 76.5%, respectively. From the conclusion obtained in this study it was evaluated that IFAT and ELISA were useful as highly specific, sensitive and stable serelogical tools for the diagnosis of canine babesiosis in Korea.

• Korean Journal of Environmental Agriculture
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• v.13 no.1
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• pp.76-82
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• 1994

A competitive indirect enzyme-linked immunosorbent assay(ELISA) was developed to detect and quantify levels of the fungicide metalaxyl in crops. Antiserum against metalaxyl was demonstrated in rabbits immunized with metalaxyl-human serum albumin(HSA) conjugate. Metalaxyl-protein conjugate was prepared by mixed anhydride and peptide coupling method with EDC. In this assay, metalaxyl-ovalbumin(OA) was $coated(8{\mu}g/ml)$ on the microtiter plate, which was incubated for 1 hr at $4^{\circ}C$ or 4 hr at $37^{\circ}C$ with diluted antiserum(1:2,000). The optimum volume ratio of antigen and antibody mixture was 0.5: 1, which was incubated for 1 hr at $20^{\circ}C$. The detection of metalaxyl bound on the surface of wells was determined by the reaction(30 min) of antirabbit Ig G-peroxidase conjugate with its substrate.

• Microbiology and Biotechnology Letters
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• v.20 no.3
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• pp.355-361
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• 1992

In order to evaluate an enzyme-linked immunosorbent assay(ELISA) for practical use in detecting aflatoxin $B_1(AFB_1)$ from cereals, we compared $AFB_1$ concentrations of samples contaminated artificially or naturally that were quantitated by the ELISA with those spiked or quantitated by HPLC. Cotton seed meals(19 items), rape seed meals(ll), soybean meals(9), and corns(3) imported from foreign countries were used as sample cereals. The standard curves of each cereal class showed that 1-100 ng/g of $AFB_1$ from cereals could be assayed by the ELISA. When artificially contaminated cereals were assayed by ELISA, the average recovery of AFB! from samples spiked to 3 ng/g and more was 138%(68-193%), although that spiked to 1 ng/g was somewhat high(268%). The average C.V. of recovery was 7.0%(0-22.2%). When naturally contaminated cereals were assayed, the concentrations of $AFB_1$ below 10 ng/g especially from rape seed meals quantitated by ELISA were much lower than those determined by HPLC. However, the concentrations of 10 ng/g and more from samples, except a few extraordinary samples. quantitated by ELISA were similar to those determined by HPLC, especially in case of cotton seed meals whose average recovery (ELISA/HPLC) was 153%. In conclusion, the ELISA was elucidated such as a practical tool to detect $AFB_1$ of 10 ng/g and more from cereals.

• Korean Journal of Veterinary Service
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• v.35 no.4
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• pp.269-273
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• 2012

A confirmatory serological test, the standard tube agglutination test (STAT) is evaluated for the diagnostic efficiency in brucellosis Korea. A total of 345 bovine samples were collected from regional veterinary branch under national brucellosis monitoring program from January 2010 to June 2012 in Korea. These samples were diagnosed as suspected serum and brucellosis positive by the Rose Bengal test (RBT) and the STAT, respectively. The STAT was compared and evaluated with three serological test such as the indirect-enzyme linked immunosorbent assay (I-ELISA), competitive-enzyme linked immunosorbent assay (C-ELISA) and fluorescence polarisation assay (FPA) prescribed for international trade by OIE. Among the 345 bovine serum samples, 302 (87.5%) were diagnosed as positive in the STAT, while 215 (62.3%), 223 (64.6%) and 194 (56.2%) serum samples were diagnosed as positive for brucellosis in the I-ELISA, C-ELISA and FPA, respectively. The STAT showed quite high positive results as compared with three prescribed tests of OIE. FPA, I-ELISA and C-ELISA have shown 60.6%, 64.9% and 67.2% correlation, respectively as compared to the STAT. However correlations of three prescribed tests ranged high 84.1~97.7%. Especially, correlation between I-ELISA and C-ELISA is quite high, 97.7%. These results suggest that the STAT has shown many false-positive reactions. Therefore, additional serological test, such as ELISAs and FPA, would be necessary to adopt as a confirmatory test in the national surveillance program of bovine brucellosis in Korea.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.2 no.3
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• pp.460-461
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• 1989

• Journal of Food Hygiene and Safety
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• v.33 no.3
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• pp.185-192
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• 2018

The purpose of this study was to develop an indirect enzyme-linked immunosorbent assay (indirect ELISA) based on a monoclonal antibody (MAb) that is specific to mackerel thermal stable-soluble protein (TSSP), that can be used for the rapid detection of mackerel in processed marine foods. Among the four MAbs (3A5-1, 2, 9, and 12) developed in previous studies, the 3A5-2 MAb that showed high specificity and sensitivity were selected and used to develop the indirect ELISA method. The detection range of the indirect ELISA was 0.02%-0.001% and the detection limit of 0.001% was shown. No cross-reaction to other marine products and food ingredients was observed by the indirect ELISA. Processed marine foods containing mackerel with ${\geq}0.3$ O.D. value at 405 nm were estimated as positive samples by the indirect ELISA. Therefore, the indirect ELISA can be used as a rapid and sensitive method to identify mackerel authenticity and adulteration in processed marine foods.