• Food Science of Animal Resources
• /
• v.43 no.6
• /
• pp.989-1001
• /
• 2023

Processed foods containing pork fat tissue to improve flavor and gain economic benefit may cause severe issues for Muslims, Jews, and vegetarians. This study aimed to develop an indirect enzyme-linked immunosorbent assay (iELISA) based on a monoclonal antibody specific to thermal stable-soluble protein in pork fat tissue and apply it to detect pork fat tissue in heat-processed (autoclave, steam, roast, and fry) beef meatballs. To develop a sensitive iELISA, the optimal sample pre-cooking time, coating conditions, primary and secondary dilution time, and various buffer systems were tested. The change in the iELISA sensitivity with different 96-well microtiter microplates was confirmed. The detection limit of iELISA performed with an appropriate microplate was 0.015% (w/w) pork fat in raw and heat-treated beef. No cross-reactions to other meats or fats were shown. These results mean that the iELISA can be used as an analytical method to detect trace amounts of pork fat mixed in beef.

• Korean Journal of Food Science and Technology
• /
• v.28 no.5
• /
• pp.953-958
• /
• 1996

A simple and rapid ELISA (enzyme-linked immunosorbent assay) system for fumonisins, a group of potentent carcinogen, was developed. To produce anti-fumonisin B1 (FB1) antibodies, FB1 conjugated to keyhole lympet hemocyanin (KLH) and Freund's adjuvant were immunized into rabbits subcutaneously 3 times. From one of the antisera showing high titer and good competition with the toxin in ELISA, polyclonal antibodies were purified. The cross-reactivities of the antibodies against fumonisin $B_1,\;B_2\;and\;B_3$ were 100%, 69%, and 166%, respectively. When competitive direct ELISA established by use of the antibody was applied to the spike test of $FB_1$ onto uncontaminated corns, the assay recovery was unstable unless 75% methanol extracts of corn were diluted to 1/100 with buffer. In that condition the mean ELISA recovery of FB1 from corns spiked $1-30\;{\mu}g/g$ was 67% and stable (coefficient of variation (CV) of each recovery percentage, 3.4%). The results suggest that the ELISA system established in this study needs no cleanup procedure and therefore would be powerful to screen a large number of corn samples contaminated with fumonisins.

• Microbiology and Biotechnology Letters
• /
• v.24 no.1
• /
• pp.119-125
• /
• 1996

In order to develop enzyme-linked immunosorbent assay (ELISA) for fumonisins, production of specific antibodies, establishment of ELISA conditions, and quantitation of the toxin from spiked corns by ELISA were performed. Fumonisin $B_1(FB_1)$ conjugated to cholera toxin (CT) with or without Freund's adjuvant was subcutaneously injected into 2 groups of rabbits. When the titer of the antisera produced by each rabbit was tested, higher titer was observed in case of the immunization with the adjuvant. By use of the antiserum showing the highest titer (1:16,000) and its purified antibodies, competitive indirect and direct ELISA's (ciELISA and cdELISA) were established, respectively. When the cross-reactivity of the antibody against fumonisin analogs was investigated by the ciELISA, it was very low against $B_3$ (2%) but high against fumonisin $B_2$ (179%). The sensitivity of the ELISAs was also very high, because the detection limit for $FB_1$ was 0.03 ppb in ciELISA and 0.3 ppb in cdELISA. When the ELISA's were applied to the spiked corns after extraction with 75% methanol, the assay recovery of $FB_1$ was too unstable to assay. However, when cleanup by strong anion exchange (SAX) cartridge was introduced to remove interfering materials, the mean ELISA recovery of $FB_1$ from corns spiked to 3~10 ppm was found to be 34.0% and stable (mean of CV, 8.2%).

• Korean Journal of Food Science and Technology
• /
• v.32 no.3
• /
• pp.564-569
• /
• 2000

An enzyme-linked immunosorbent assay(ELISA) was developed for the detection of milk proteins in processed foods. The ${\alpha}_{s1}-casein({\alpha}_{s1}-CN)$, a heat stable major milk protein, was immunized into rabbits to produce specific antibodies. When competitive indirect ELISA(ciELISA) using $anti-{\alpha}_{s1}-CN$ antibodies was established, its detection limit was $0.1\;{\mu}g/mL$. The reactivities of the specific antibodies toward ${\alpha}_{s1}-CN$, skim milk, ${\beta}-CN$ and whey protein isolate(WPI) were 100, 37, 0.14 and 0.04%, respectively, as determined by ciELISA. However $anti-{\alpha}_{s1}-CN$ antibodies did not have any reactivity to other milk proteins such as ${\beta}-lactoglobulin,\;{\alpha}-lactalbumin$, bovine serum albumin, and isolated soy protein. When sandwich ELISA was established, its detection limit was $0.01\;{\mu}g/mL$ which was 10 times more sensitive than that of ciELISA. In the spike test which was performed by adding 1-10% of whole CN to market milk, mean assay recovery as determined by sandwich ELISA was 94.8%(CV, 8.2%). Food stuffs and dairy products were assayed by sandwich ELISA to show 29, 0.13, 0.25, and 6.9% of whole CN in skim milk powder, WPI, semi-solid yoghurt, and processed cheese, respectively.

• Korean Journal of Food Science and Technology
• /
• v.28 no.6
• /
• pp.1184-1187
• /
• 1996

A simple and rapid detection system for $aflatoxin\;M_1\;(AFM_1)$ in cow's milk by an enzyme-linked immunosorbent assay (ELISA) was developed. Specific antibodies against $AFM_1$, conjugated to bovine serum albumin $(AFM_1-BSA)$ were raised in rabbits and purified. The cross-reactivities of the antibodies against aflatoxin analogs were less than 29.9%. When a competitive direct ELISA (cdELISA) for $AFM_1$, established by use of the antibodies was applied to the spike test of $AFM_1$ onto uncontaminated cow's milk, the assay recovery was unstable unless cow's milk was diluted to 40% (2:3) with phosphate buffered saline (PBS). In that condition of sample dilution, the mean ELISA recovery of $AFM_1$, from the cow's milk was 113% (coefficient of variation (CV) of each recovery percentage, 8.2%) in the range of $0.3{\sim}3.0\;ppb$. These results showed that the ELISA system could be a convenient tool to monitor the contamination of AFM1 more than 0.5 ppb in cow's milk (FDA allowance limit) easily.

• Korean Journal of Food Science and Technology
• /
• v.44 no.1
• /
• pp.1-7
• /
• 2012

There have been few studies related to ELISA for mackerel. In this study we developed a sandwich ELISA for mackerel in processed foods using rabbit polyclonal antibodies against mackerel parvalbumin, the major allergen of mackerel and heat-stable protein. The parvalbumin was purified by ammonium sulfate precipitation and Sephadex G-50 column chromatography. From the standard ELISA curves, the detection limit of parvalbumin was 3 ng/mL and the detection range of mackerel was 5-5,000 ${mu}g/mL$. We further investigated the cross-reactivity of the antibodies toward mackerel, mackerel pike, salmon, flatfish, armorclad rockfish, cod fish, squid, shrimp, blue crab, and lobster. The antibodies were specific for mackerel only. The mean assay recoveries in cooked cream soup, baby food, sausage, and sauce spiked with 0.01 to 0.3% mackerel were 104, 101, 54, and 0%, respectively. In sample tests of 16 commercial items, the qualitative coincidence ratio of assay result and indication was 75%.

• Korean Journal of Veterinary Service
• /
• v.19 no.1
• /
• pp.30-38
• /
• 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).

• Parasites, Hosts and Diseases
• /
• v.21 no.2
• /
• pp.270-280
• /
• 1983

Agar-gel Diffusion test (AGD), Counterimmunoelectrophoresis(CIEP) and Enzyme-Linked Immunosorbent Assay(ELISA) were examined with the sera of skin test positives for paragonimiasis. The crude antigen (Paragcnimus whole worm extracts: protein concentration, 7.56mg/m1) and human sera were used in AGD and CIEP. And in ELISA test, diluted antigen with 1:40, 000 of crude antigen and diluted sera with 1:100, 1:200 were used in the test. The positive identical ratio between AGD and CIEP reactions is 985 and negative identical ratio is 100%. One or three precipitin bands are observed in AGD. One to seven precipitin bands are also revealed in CIEP. Especially, deeply stained bands are observed in CIEP than those of AGD. The positive identical ratios between AGD and ELISA tests are 96% in 1:100 diluted sera, and 94% in 1:200 diluted sera. But the negative identical ratios between AGD and ELISA tests are 97% and 99% respectively in 1:100 and 1:200 diluted sera. The positive identical ratios between CIEP and ELIEA tests are 98% and 96% respectively in 1:100 and 1:200 diluted sera, but also 97% and 99% in 1:100 and 1:200. Control sera, such as clonorchiasis, amoebiasis and toxoplasmcsis, revealed all negatives with Paragonimus antigen in AGD, CIEP and ELISA tests. By above results, ELISA was mcst sensitive, next CIEP and AGD, But AGD test apprars to be more useful when used to crude antigen without cross rfacticn with other parasitic infections. CIEP test is basically equal in terms of precipitin reaction, but CIEP is able to be detected more sensitively and rapidly though less simple in handiwork than AGD. Consequently, three methods for inlmunological tests of paragonimiasis have gccd correlations with one another. Also, each of these has both merits and demerits in iymunolcgical test for paragonimiasis. But the ELISA test was proved to be the most sensitive and convenient tool for mass screening test, especially in cacti of using purified antigen.

• Clinical and Experimental Reproductive Medicine
• /
• v.17 no.1
• /
• pp.71-80
• /
• 1990

New immunoassay systems for the detection of anti-sperm antibodies were developed. For this, sperm surface protein was purified by the immunoaffinity column prepared by the coupling of rabbit anti-human IgG antibodies to Sepharose-4B. Fraction eluted by tris-HCI buffer containing SDS showed a single band having molecular weight of about 60KD on electrophoresis. Enzyme HRP labelled goat anti-human IgG and chemiluminescence aminobutylethyl-isoluminol(ABEI) labelled rabbit anti-human IgG were used for ELISA and CIA, respectively. These two labelled conjugate bound well with human IgG. When serum dilution curves were made to titrate positive serums, two kinds of curves with steep and sluggish slopes were obtained Serum samples were categorized into 3 groups: positive, weak positive and negative based on slope of curve and O.D. values at 1:160 dilution of serum. When ELISA and CIA were compared to conventional method Kibrick test by the determinations of 62 male serums with different diagnosis, the results of ELISA and CIA agreed well, but both disagreed with that of Kibrick test. This study showed that purified sperm surface antigen can be used to develope solid-phase immunoassay systems such as ELISA and CIA which may eliminate the problems encounted the immobilization of living sperm in other tests.

• Journal of fish pathology
• /
• v.10 no.2
• /
• pp.75-86
• /
• 1997

Solid phase enzyme linked immunosorbent assay (ELISA) was developed to detect the whole cells of Edwardsiella tarda from infected tissues of flounder. Cross-reaction test was performed by ELISA against fish pathogens such as A. hydrophila ATCC7966. V. anguillarum HYUFP5001, Y, ruckeri 11-4, E. ictaluri and Streptococcus sp. NG8206. Rabbit anti-E, tarda Edk-2 sera highly cross-reacted with A. hydrophila ATCC7966 and V. anguillarum HUFP5001. However, the cross-reaction was removed by using the anti-serum pre-adsorbed with A, hydrophila ATCC7966 FKC. The intra-species cross-reaction among E. tarda isolates was very high. ELISA with the whole cell antigens present in tissue homogenate appeared with highly decreased sensitivity, presumably by the co-coating of lipid or proteins in tissues. Thus, it would be necessary to use the infected tissue homogenates diluted more than 100 times with PBS for diagnosis. Interestingly, compared with the using of FKC antigen, the direct detection of viable cells in tissue homogenate showed more sensitive results with detection limit of $1{\times}10^3$ cells/ml in buffer or diluted tissue homogenate. Consequently, the ELISA method developed in this study was specific, rapid and sensitive for diagnosing edwardsiellosis.