• Korean Journal of Food Science and Technology
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• v.36 no.5
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• pp.828-832
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• 2004

Enzyme-linked immune sorbent assay (ELISA) was applied to quantify soy protein in fresh soybean curd (bean curd) produced by combination of genetically modified (GM) and genetically not modified (non-GM) soybeans. Antibodies against 113 and 24 kDa proteins, which appeared only in non-GM bean curd (specific band), and in both non-GM and GM bean curds (non-specific band) based on SDS-PAGE results, were prepared by immunization to rabbit. Through ELISA using either antibody, GM bean curd protein content was determined at dilution rates of $10^{-1}-10^{-6}$. Standard curve showing relationship between ELISA optical density and non-GM protein content was produced using antibody against 113 kDa protein at protein dilution between $10^{-7}\;to\;10^{-6}$, highly antigen content-dependent dilution. Bean curd prepared by random combinations of GM and non-GM soybeans were analyzed by ELISA, and standard curve was produced. Results reveal non-GM protein content of bean curd could be quantified with higher than 93% accuracy.

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

• Journal of Food Hygiene and Safety
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• v.9 no.3
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• pp.123-131
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• 1994

An enzyme-linked immunosorbent assay(ELISA) was developed for the detection of residual gentamicin(GM) in edible animal products. The immunogen(GM-KLH conjugate) and coating antigen(GM-BSA conjugate) were prepared by coupling GM sulfate to keyhole limpet hemocyanin(KLH) and bovine serum albumin(BSA) in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, respectively. Polyclonal antibody to GM was produced in rabbits(New Zealand White, female) by using the immunogen and the antibody titer was measured by indirect ELISA. A competitive ELISA was developed using GM-bovine serum albumin conjugate as a coating antigen, GM(as standards or sample), polyclonal antibody to GM, secondary antibody conjugated with horseradish peroxidase as an enzyme, and H2O2 and o-phenylenediamine dihydrochloride as a substrate and a chromophore, respectively. The detection limit of GM was 10 ng/ml and the standard curve of GM(n=26) was linear up to 10 $\mu\textrm{g}$/ml in this competitive ELISA system. There were no cross-reactivities of the partially purified antibody between GM and the various antibiotice such as amikacin, benzyl-penicillin, chloramphenicol, erythromycin, furazlidone, kanamycin, neomycin, oleandomycin, streptomycin, sulfathiazole and thiamphenicol(CR50<0.05%)

• Journal of Plant Biotechnology
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• v.30 no.1
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• pp.97-102
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• 2003

Lettuce (Lactuca sativa) was transformed with Agrobacterium tumefacience LBA4404 containing human granulocyte macrophage colony stimulating factor (hGM-CSF) gene to produce in cell suspension cultures. Cell suspension culture was established using callus from transgenic lettuce plant. Integration of hGM-CSF gene into plant chromosome was confirmed through genomic PCR and Southern blot analysis. In addition, Northern blot analysis indicated the expression of the introduced hGM-CSF gene in transformed lettuce. The recombinant hGM-CSF was expressed in transgenic cell cultures derived from transgenic plants as a yield of about 149.0 $\mu\textrm{g}$/L in culture filtrate, which was determined by ELISA. These results demonstrated that transformed lettuce cell suspension cultures could be used as a production system of therapeutic proteins such as hGM-CSF.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.5
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• pp.1397-1403
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• 2004

By recently study, GM (Gamipaemo-tang) treatment have worked well on the allergic asthma. The purpose of this study was effect of GM extract on helper T cell, major regulator of immune system. Splenic cells from 8-week BALB/c mice were cultured in GM containing media without activation for 48 hours. The MTS assay and flow cytometry study revealed that lymphocyte treated with GM were not effective on CD4+ T cells. Subsequently CD4+ T cells were isolated and cultured in GM containing media. Either GM were not effective on CD4+ T cell without APCs. By FACS scan analysis, the expression of INF-γ, IL-4 were down-regulated in the condition skewed Th1 and Th2 cells respectively, Using ELISA analysis, the expression of INF-γ is up-regulated and IL-4 is down-regulated in the condition skewed Th1, Th2 cells respectively. With RT-PCR analysis, the expression of mRNA for INF-γ is down-regulated and IL-4 is down-regulated in the condition skewed Th1 and Th2 cells respectively. The result suggests that GM inhibited the differetiation of Th2 cells significantly and indicates GM could enhance anti-allergic immune system.

• Korean Journal of Environmental Agriculture
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• v.32 no.3
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• pp.231-239
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• 2013

BACKGROUND: The disease resistant (OsCK1) rice was generated by inserting choline kinase (CK1) and phosphinothricin acetyltransferase (PAT) genes isolated from Oriza sativa and Streptomyces hygroscopicus into the genome of rice (Nakdongbyeo). With the potential problems of safety, the non-target organism evaluation is required as an essential element for the environmental risk assessment of genetically modified (GM) crops. In present study, we studied the effects on survival of Misgurnus anguillicaudatus and Cyprinus carpio, commonly used as a model organism in ecotoxicological studies. METHODS AND RESULTS: The M. anguillicaudatus and C. carpio were fed on disease resistant (OsCK1) rice and non-genetically modified (non-GM) rice (Nakdongbyeo) to 0, 10, 100, 1,000 and 5,000 mg/L, as treatment concentration respectively. The OsCK1 rice used for the test was confirmed to have the OsCK1/PAT gene expression by the PCR and ELISA analysis. Feeding test showed that no significant differences in cumulative immobility and abnormal response of M. anguillicaudatus and C. carpio fed on between OsCK1 rice and non-GM rice. The 96hr-$LC_{50}$ values showed no difference between OsCK1 rice (>5,000 mg/L) and non-GM rice (>5,000 mg/L). CONCLUSION(S): The results of this study suggested that there was no significant difference in toxicity for M. anguillicaudatus and C. carpio between OsCK1 rice and non-GM counterparts.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.2
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• pp.135-141
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• 2003

The human granulocyte-macrophage colony stimulating factor (hGM-CSF) gene was introduced into tobacco plants. The cell suspension culture was established from leaf-derived calli of the transgenic tobacco plants in order to express and secrete a biologically active hGM -CSF. The recombinant hGM-CSF from the transgenic plant cell culture (prhGM-CSF) was identified as a yield of about 180 ${\mu}$g/L in the culture filtrate, as determined by ELISA. The addition of 0.5 g/L polyvinylpyrrolidone (PVP) to the plant cell culture medium both stabilized the secreted prhGM-CSF and increased the level of production approximately 1.5-fold to 270 ${\mu}$g/L. The biological activity of the prhGM-CSF was confirmed by measuring the proliferation of the hGM-CSF-dependent cell line, TF-1. Interestingly, the specific activity of the prhGM-CSF was estimated to be approximately 2.7 times higher than that of a commercially available preparation from E. coli.

• Journal of Microbiology and Biotechnology
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• v.10 no.3
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• pp.287-292
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• 2000

Recombinant Aspergillus niger was constructed to express and secrete a biologically active murine granulaocyte macrophage-colony stimulating factor (mGM-CSF). A 500 bp fragment encoding the signal peptide and terminator of glyceraldehyde-3-phosphate dehydrogenase (gpd). The hygromycin phosphotrasferase gene (hph) was used as a selection marker for the fungal transformants. An expression vector was introduced into A. niger ATCC 9642, and a Northern blot analysis indicated the presence of a considerable amount of transcripts from the introduced mGM-CSF. The biological activity of recombinant mGM-CSF (rmGM-CSF) isolated from the culture filtrate was confirmend by measuring the proliferationof the GM-CSF dependent FDC-P1 cell line. It appeared that rmGM-CSF was amenable to the proteolytic activity produced by A. niger, since biological actibity was only observed when the transformants were grown in a protease-repressing medium, and the activity of rmGM-CSF dramatically decreased with an increase of age of the culture. The yield of rmGM-CSF, as determined by ELISA. was 640 ng/l of culture filtrate. Accordingly, its specific activity is estimated to be approximately two-and-a-half times higher than that of a commercial preparation from E. coli.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.206-209
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• 2001

Recombinant human GM -CSF was expressed and secreted from transgenic rice cell suspension cultures in its biologically active form. This was accomplished by transforming rice callus tissues with an expression vector, pMYN44. containing the hGM -CSF cDNA. Regulated expression and secretion of hGM -CSF from this vector achieved using the promoter, signal peptide, and terminator from a rice alfa-amylase gene Amy3D. The Amy3D gene is expressed in response to sugar deprivation. The recombinant hGM -CSF was expressed from the transgenic rice cell culture on the sugar-free medium as a yield of about 110 mg/L in the culture filtrate, which was determined by ELISA. Biological activity of hGM-CSF was confirmed by measuring the proliferation of the hGM -CSF dependent TF -1 cells.(This work was supported by a grant from the NRL program of the Korean Ministry of Science and Technology. Shin, Y.- J.. Lee. J.-H and Kwon, T.-H. have been supported by BK21 program from the Korean Ministry of Education)

• Journal of Food Hygiene and Safety
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• v.33 no.3
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• pp.193-199
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• 2018

In this study, PAT protein of genetically modified maize was prepared from the recombinant E. coli strain BL21 (DE3), and mice were immunized with the recombinant PAT protein. After cell fusion and cloning, two hybridoma cells (PATmAb-7 and PATmAb-12) were chosen since the monoclonal antibodies (Mabs) produced by them were confirmed to be specific to PAT protein in the indirect enzyme-linked immunsorbent assay (ELISA) and western blot tests. There were no cross-reactions of either Mabs to other GM proteins or to the extracts of non-GM maize. The ELISA based on the PATmAb-7 can sensitively detect 0.3 ng/g PAT protein in corn. These results indicate that the developed Mabs can be used as bio-receptors in the development of immunosensors and biosensors for the rapid and simple detection of GM corn adulterated in foods.