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http://dx.doi.org/10.13103/JFHS.2018.33.3.193

Production and Characterization of Monoclonal Antibodies Specific to PAT Protein Expressed in Genetically Modified Herbicide-Resistance Maize  

Kim, Sol-A (Division of Applied Life Science, Graduate School, Gyeongsang National University)
Lee, Jeong-Eun (Division of Applied Life Science, Graduate School, Gyeongsang National University)
Shim, Won-Bo (Department of Agricultural Chemistry and Food Science & Technology, Gyeongsang National University)
Kang, Sung-Jo (MADI-LAB, Inc.)
Chung, Duck-Hwa (Department of Agricultural Chemistry and Food Science & Technology, Gyeongsang National University)
Publication Information
Journal of Food Hygiene and Safety / v.33, no.3, 2018 , pp. 193-199 More about this Journal
Abstract
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.
Keywords
GM maize; PAT protein; Hybridoma cell; Monoclonal antibody;
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1 Bonny, S.: Genetically modied herbicide-tolerant crops, weeds, and herbicides: Overview and impact. Environ Manag., 57, 31-48 (2016).   DOI
2 Chung, K.H.: Genetically Modified Organisms. Health and Welfare Forum, 64-73 (2011).
3 Shin, W.S., Kim, M.H.: Optimized condition of genomic DNA extraction and PCR methods for GMO detection in potato. Korean J. Food Sci. Technol., 35, 591-597 (2003).
4 Available from http://foodsafetykorea.go.kr/portal/board/board.do/, Accessed Aug. 29 (2017).
5 Zhu, H.J., Chen, Y., Li, Y.H., Wang, J.M., Ding, J.T., Chen, X.P., Peng, Y.F.: A 90day safety assessment of genetically modified rice expressing Cry1Ab/1Ac protein using an aquatic animal model. J. Agric. Food Chem., 63, 3627-3633 (2015).   DOI
6 Zou, S., Huang, K., Xu, W., Luo, Y., He, X.: Safety assessment of lepidopteran insect-protected transgenic rice with cry2A gene. Transgenic Res., 25, 163-172 (2016).   DOI
7 Hooftman, D.A.P., Bullock, J.M., Morley, K., Lamb, C., Hodgson, D.J., Bell, P., Thomas, J., Hails, R.S.: Seed bank dynamics govern persistence of Brassica hybrids in crop and natural habitats. Ann. Bot.,115, 1-11 (2014).
8 Duke, S.O., Lydon, J., Koskinen, W.C., Moorman, T.B., Chaney, R.L., Hammerschmidt, R.: Glyphosate effects on plant mineral nutrition, crop rhizosphere microbiota, and plant disease in glyphosate-resistant. Crops. J. Agric. Food Chem., 60, 10375-10397 (2012).   DOI
9 Delaney, B., Karaman, S., Roper, J., Hoban, D., Sykes, G., Mukerji. P., Frame, S.R.: Thirteen week rodent feeding study with grain from molecular stacked trait lepidopteran and coleopteran protected (DP-OO4114-3) maize. Food Chem. Toxicol., 53, 417-427 (2013).   DOI
10 Devos, Y., De Schrijver, A., De Clercq, P., Kiss, J., Romeis, J.: Bt-maize event MON 88017 expressing Cry3Bb1 does not cause harm to non-target organisms. Transgenic Res., 21, 1191-214 (2012).   DOI
11 Nilufer, D., Boyacioglu, D.: Comparative study of three different methods for the determination of aflatoxins in tahini., J. Agric. Food Chem. 50, 3375-3379 (2002).   DOI
12 Cao, S.S., Xu, W.T., Luo, Y.B., He, X.Y., Yuan, Y.F., Ran, W.J., Liang, L.X., Huang, K.L.: Metabonomics study of transgenic Bacillus thuringiensis rice (T2A-1) meal in a 90-day dietary toxicity study in rats. Mol. Bio. Syst., 7, 2304-2310 (2011).
13 Ford, C.S., Allainguillaume, J., Fu, T.R., Mitchley, J., Wilkinson, M.J.: Assessing the value of imperfect biocontainment nationally: rapeseed in the United Kingdom as an exemplar. New Phytologist., 205, 1342-1349 (2015).   DOI
14 Ford, C.S., Allainguillaume, J., Grilli-Chantler, P., Cuccato, J., Allender, C.J., Wilkinson, M.J.: Spontaneous gene flow from rapeseed (Brassica napus) to wild Brassica oleracea. Proc. Biol. Sci., 273, 3111-3115 (2006).   DOI
15 Langone, J.J., Van Vunakis, H.: Aflatoxin B; specific antibodies and their use in radioimmunoassay. J. Natl. Cancer Inst. 56, 591-595 (1976).   DOI
16 Keese, P.: Risks from GMOs due to horizontal gene transfer. Environ. Biosafety Res., 7, 123-149 (2008).   DOI
17 Kohler, G., Milstein, C.: Continuous cultures of fused cells producing antibody of predefined specificity. Nature, 256, 495-501 (1975).   DOI
18 Available from https://www.foodsafetykorea.go.kr/portal/board/boardDetail.do/, Accessed Mar. 24 (2017).
19 Laemmli, U.K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 15, 680-685 (1970).
20 Shon, D.H.: Development of immunoassay system for the detection of genetically modified soybeans. Bull. Food Technol., 15, 59-69 (2002).
21 Xu, W.T., Huang, K.L., Deng, A.K., Luo, Y.B.: Enzyme linked immunosorbent assay for PAT protein detection in genetically modified rape. Chinese J. Agric. Biotechnol., 3, 177-181 (2006).   DOI
22 Mckearn, T.J., Weiss, A., Stuart, F.P., Fitch, F.W.: Selective suppression of humoral and cell-mediated immune responses to rat alloantigens by monoclonal antibodies produced by hybridoma cell lines. Transplant Proc., 11, 932-935 (1979).
23 George, C.W., Susott, R.A.: Effects of ammonium phosphate and sulphate on the pyrolysis and combustion of cellulose. USDA Forest Service INT-90 (1971).
24 Towbin, H., Staehelin, T., Gordon, J.: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc. Natl. Acad. Sci., 76, 4350-4354 (1979).   DOI
25 Available from http://www.fao.org/fileadmin/user_upload/gmfp/docs/SYHT0H2_report.pdf/, Accessed Nov. 4 (2014).