• Korean Journal of Food Science and Technology
• /
• v.35 no.4
• /
• pp.556-560
• /
• 2003

It was determined whether the sandwich ELISA using specific anti-CP4 EPSPS polyclonal and monoclonal antibodies, developed in the previous study, could be applied to detect GM soybean or not. The soybeans (47 imported and 20 domestic soybeans) were analyzed by a sandwich ELISA. The results of imported soybeans were divided into two groups which were high contents $(39.1{\pm}13.5\;{\mu}g/g,\;n=33)$ and low contents of CP4 EPSPS $(2.6{\pm}1.2\;{\mu}g/g,\;n=14)$. The ratio of GM in imported soybeans was about 70.2%. One the other hand, the contents of CP4 EPSPS in domestic soybeans was very low $(0.9{\pm}0.5\;{\mu}g/g,\;n=20)$ which determined to be non-GM soybeans. In case of soybean sprouts, the contents of CP4 EPSPS in soybean sprouts were different between GM and non-GM soybean sprout. The CP4 EPSPS in cotyledon of GM soybeans sprout was higher than that in root hair. The contents of CP4 EPSPS in soybeans sprout of domestic soybeans were very low. Thus, it was possible to determine that the soybeans sprout was made of GM or non-GM soybeans. Also, PCR experiment showed that the sandwich ELISA was accurate to distinguish the soybeans to be GM or non-GM. These results showed the sandwich ELISA could determine the soybeans were GM or non-GM, rapidly and simply.

• Food Science and Preservation
• /
• v.12 no.1
• /
• pp.23-27
• /
• 2005

Yolk index (In, Haugh unit (BU) and weight loss of quail egg were measured to evaluate the effect of composite film (SPI/corn starch). Also, the effect of composite film was investigated to extend the shelf-life of sandwich foods. The quality characteristics of sandwich food was measured by the weight increment The weight reduction ratio for quail egg coated with composite film showed $8\%$ increment after 20 day storage. Yolk index and Haugh unit were significantly different between the uncoated and coated quail eggs with composite film solution. Sandwich coated with composite film showed the less weight increase for 12 hour storage compared to controls.

• Korean Journal of Food Science and Technology
• /
• v.35 no.3
• /
• pp.366-372
• /
• 2003

Enzyme-linked immunosorbent assay (ELISA) for assaying the 5-enolpyruvyshikimate-3-phosphate synthase from Agribacterium sp. CP4 (CP4 EPSPS) in genetically modified soybeans was developed. Polyclonal and monoclonal antibodies (Pab, Mab) specific to the CP4 EPSPS were produced. When using the Pab, the detection limit of sandwich ELISA toward CP4 EPSPS (0.03 ${\mu}g/mL$) was better than that of competitive indirect ELISA(ciELISA) (1 ${\mu}g/mL$). It was found that 2 of 3 monoclonal antibodies, Mab1 and Mab2, recognized the same antigenic determinant on CP4 EPSPS, but Mab3 recognized different antigenic determinant when competitive ELISA was performed using the Mabs. On the other hand, when the sensitivity of sandwich ELISA using combination of Pab and/or Mabs was determined, the sandiwich ELISA using Mab2 as a capture antibody and Pab-HRP as a secondary antibody showed the lowest detection limit of CP4 EPSPS (0.02 ${\mu}g/mL$). The sandwich ELISA developed in this study could be applied to detect glyphosate-tolerant soybeans.

• Food Science of Animal Resources
• /
• v.24 no.2
• /
• pp.176-181
• /
• 2004

The specificity of sandwich enzyme-linked immunosorbent assay (ELISA) to detect Salmonella in milk was determined in this study. The antibodies used in sandwich ELISA were egg yolk immunoglobulin G (IgY) obtained after immunization of hen with outer membrane protein (OMP) fraction from Salmonella typhimurium and rabbit IgG obtained after immunization of rabbit with the purified OMP with the molecular weight of 40,000. The immunoblot assay showed that the IgY reacted strongly with OMP with the molecular weight of 6,000 and the rabbit IgG reacted strongly with OMP with the molecular weights of 40,000, 35,000, and 6,000 from the bacteria including Salmonella which belongs to Enterobacteriaceae. The IgY and rabbit IgG also reacted with other proteins from Salmonella typhimurium in immunoblot assay. Competitive ELISA showed that IgY showed specifity to react with two strains of Salmonella typhimurium and Salmonella cholerasuis but not with Escherichia coli and Yersinia enterocolitica. Two strains of Salmonella typhimurium added to UHT milk showed the highest absorbance of all the bacteria used in the sandwich ELISA. Some strains of Salmonella cholerasuis showed higher absorbances than non-Salmonella bacteria.

• Journal of Microbiology and Biotechnology
• /
• v.13 no.5
• /
• pp.794-799
• /
• 2003

Enzyme-linked immunosorbent assay (ELISA) was developed for the rapid detection of Fusarium species, known as harmful fungi in food. One of the hybridoma cell lines (lB8) which produced a monoclonal antibody (Mab) specific to Fusarium extracellular polysaccharide (EPS) was screened and the Mab was produced and purified. A detection limit of the sandwich ELISA against F. moniliforme EPS was $0.001\;\mu\textrm{g}/ml$ in the standard curve. Among the 59 strains tested, most Fusarium species showed hight reactivity with Mab lB8, even when the culture broths were diluted 100,000 times. On the other hand, the other genera, except A. versicolor and Trichoderma viride, had no reactivity. When 1 to $100\;\mu\textrm{g}$ of F. moniliforme EPS was spiked into rice, potato, and mandarine orange, the average recoveries were 151%, 84%, and 94%, respectively, determined by sandwich ELISA. The correlation coefficients between the EPS levels determined by sandwich ELISA and the dry mycelial weight of the liquid culture of F. moniliforme, as well as between the EPS and colony forming unit in solid culture of potato, were 0.97 and 0.91, respectively.

• Journal of Microbiology and Biotechnology
• /
• v.9 no.6
• /
• pp.764-772
• /
• 1999

To develop an enzyme-linked immunosorbent assay (ELISA) for detecting mold, we produced anti-mold polyclonal antibodies by immunizing extracellular polysaccharide (EPS) of Aspergillus flavus or Penicillium citrinum in rabbits subcutaneously. Using the purified antibody (Ab) and Ab-HRP conjugate, a sandwich ELISA for EPS was established. The standard curve of the ELISA showed the detection limit for P citrinum EPS to be $0.003{\;}\mu\textrm{g}/ml$. The cross-reactivities of the anti-P citrinum EPS Ab toward components of P citrinum such as EPS, liquid, and solid culture mycelium were 100, 10.5, and 0.58%, respectively, and those toward components of A. flavus such as EPS, liquid and solid culture mycelium, and spore were 300, 0.67, 0.29, and 0%, respectively. When the reactivities toward culture broths of 59 mold strains were tested by the sandwich ELISA, most of the Aspergillus (16 of 18) and Penicillium (14 of 16) strains along with one of the two Cladosporium strains gave positive signals in the culture broths even when diluted 1,000 fold, while the rest of species such as Fusarium, Absidia, Alternaria, and Candida gave negative signals. When the water extracts of 30 corn samples were analyzed by the sandwich ELISA, the EPS in the com could be detected in the concentration range of $0.1-1.6{\;}\mu\textrm{g}/g$.

• Journal of Microbiology and Biotechnology
• /
• v.14 no.2
• /
• pp.385-389
• /
• 2004

The antibody-mix sandwich enzyme-linked immunosorbent assay (Ab-mix sELlS A) system was developed in order to simultaneously detect the extracellular polysaccharide (FPS) of Aspergillus, Penicillium, or Fusarium species using one detection system. The detection limit and detection range of the Ab-mix sELISA towards EPS of Penicilliun citrinum were not changed, and those towards Fusarium moniliforme EPS were changed a little compared to that of individual sandwich ELISA [9, 10]. The fungal culture filtrates of Aspergillus and Penicillium species showed nearly similar reactivity towards Ab-mix sELISA as that of sELISA using the MAb lB8 alone [9]. Also, the fungal culture filtrates of Fusarium species showed nearly the same reactivity towards Ab-mix sELISA as that of sELISA using the MAb lB8 alone [10]. Thus, this ELISA system showed that the three genera of molds, Aspergillus, Penicillium, or Fusarium, which are three major important molds producing mycotoxins in food or agricultural commodities, could be detected at the same time, using one detection system.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.30 no.4
• /
• pp.600-604
• /
• 2001

Immunoassay was used to study the detection method of irradiated shrimp. Sandwich ELISA was formatted with monoclonal antibody (Ab) (M-IgG) and polyclonal Abs (P-IgG) and polyclonal Abs (P-IgG) individually produced against brown shrimp tropomyosin (TPM) as an antigen. When M-IgG was used as a coating Ab to capture TPM, and P-IgG were used as reaction Ab against captured TPM could be detected in the range of 12.5 to 50 $\mu\textrm{g}$/mL. Detected concentrations of TPM from irradiated shrimp decreased dose-dependently, and the concentration of Ag by combination of irradiation with heating or freezing treatments also decreased. This results suggests the possibility for Sandwich ELISA, one of immunological analyses, to be applied for detecting irradiated shrimp.

• Food Science and Biotechnology
• /
• v.15 no.1
• /
• pp.133-137
• /
• 2006

Wheat gluten-chitosan composite film (WGCCF) can prevent moisture migration and enhance the antimicrobial properties of gluten in intermediate-moisture foods like sandwiches. To mimic the structure of actual sandwich-type products we developed multi-layer food models, where moisture content and water activity differ. Water activity gradients direct moisture migration and therefore determine product characteristics and product stability. A 10% wheat gluten film-forming solution was mixed with chitosan film-forming solution (0-3%, w/w) and evaporated to generate WGCCF. Addition of 3% chitosan enhanced the mechanical properties of the film composite, lowered its water vapor permeability, and improved its ability to protect against both, Streptococcus faecalis and Escherichia coli, in a 24 hr sandwich test (reduction of 1.3 and 2.7 log cycles, respectively, compared to controls). Best barrier and antimicrobial performance was found for 3% chitosan WGCCF at pH 5.1. Film of this type may find application as barrier film for intermediate-moisture foods.

• Food Science of Animal Resources
• /
• v.18 no.1
• /
• pp.27-34
• /
• 1998

This study was carried out to evaluate of the microbial and sensory quality of ready-made hamburger and sandwich. Initial total plate count of hamburger for establishment A and B were 1.2$\times$102 cfu / g and 3.4$\times$102 cfu / g, respectively, and for establishment C was 7.9$\times$104 cfu / g. After 48 hour storage at 1$0^{\circ}C$, total plate count of hamburger for establishment A and B increased to 1.2$\times$104 cfu / g and 6.8$\times$103, respectively, and for establishment C increased to 1.2$\times$107 cfu / g. Initial total plate count of sandwich for establishment A and B were 3.2$\times$102 cfu / g 7.9$\times$102 cfu / g, respectively, and for establishment C was 1.1$\times$105 cfu / g. After 48 hour storage at 1$0^{\circ}C$, total plate count of hamburger for establishment A and B increased to 8.1$\times$103 cfu / g and 2.3$\times$104, respectively, and for establishment C increased to 4.4$\times$108 cfu / g. No E. coli, Salmonella, Vibrio, and Staphylococcus aureus were detected under simulated storage conditions. There was no significant changes in pH, acid value, and volatile nitrogen number under simulated conditions. In sensory evaluation of hamburger and sandwich, sensory score was lowered by increase of total plate count.