|
OALib Journal期刊
ISSN: 2333-9721
费用：99美元
|
|
|
|
噻虫嗪单克隆抗体制备及酶联免疫吸附分析方法的建立
DOI: 10.3724/SP.J.1096.2012.11245, PP. 1098-1103
Keywords: 噻虫嗪,半抗原,单克隆抗体,残留,酶联免疫吸附法
Abstract:
通过化学修饰合成了噻虫嗪人工半抗原,采用碳二亚胺法将该半抗原与牛血清蛋白(BSA)和卵清蛋白(OVA)偶联,成功制备了分子结合比合理的免疫原和包被原。经过免疫原免疫6周龄Balb/c小鼠、PEG介导免疫鼠脾细胞和骨髓瘤细胞的融合和阳性杂交瘤细胞的筛选和克隆化,获得了效价高达1:6.4×105的抗噻虫嗪单克隆抗体,抗体亚类为IgG1型。优化了ELISA实验条件,建立了基于单克隆抗体技术的噻虫嗪残留间接竞争ELISA方法。本方法的抑制中浓度(IC50)为0.0255mg/L,检测灵敏度(IC20)为0.0022mg/L,检出限(IC10)为0.001mg/L。除噻虫胺外,该抗体与其它噻虫嗪结构类似物无交叉反应。以自来水为基质的噻虫嗪添加回收实验显示,0.01,0.5和10.0mg/L添加水平的回收率均大于75%,且各添加水平重复测定8次的相对标准偏差均小于8%,说明所建立的间接竞争ELISA准确度高,重复性好,适合水中噻虫嗪残留的检测。
| [1] | 1 Maienfisch P, Huerlimarm H, Rindlisbacher. Gsell L, Dettwiler H, Haettenschwiler J, Sieger E, Walti M. Pest Manag. Sci., 2001, 57(2): 165~176
|
| [2] | 2 Tan J G, Salgado V L, Hollingworth R M. Pest Manag. Sci., 2008, 64(1): 37~47
|
| [3] | 张国生,侯广新. 农药科学与管理, 2004, 25(3): 22~26
|
| [4] | 5 Ensley S. In Veterinary Toxicology: Basic and Clinical Principles, R. Gupta(Ed.), Elsevier, Amsterdam, The Netherlands, 2007: 505~507
|
| [5] | 6 Mota-Sanchez D, Hollingworth R M, Grafius E J, Moyer D D. Pest Manag. Sci., 2006, 62(1): 30~37
|
| [6] | 7 Obana H, Okihashi M, Akutsu K, Kitagawa Y, Hori S. Agric. Food Chem., 2003, 51(9): 2501~2501
|
| [7] | 8 Di Muccio A, Fidente P, Barbini D A, Dommarco R, Seccia S, Morrica P. J. Chromatogr. A, 2006, 1108(1): 1~6
|
| [8] | 9 Venkateswarlu P, Rama Mohan K, Ravi Kumar Ch, Seshaiah K. Food Chem., 2007, 105(4): 1760~1766
|
| [9] | 11 García-Chao M, Agruna MJ, Flores Calvete G, Sakkas V, Llompart M, Dagnac T. Anal. Chim. Acta, 2010, 672(1-2): 107~113
|
| [10] | 16 Gosling J P. Immunoassays a Practical Approach, America: Oxford University Press, 2000: 41
|
| [11] | 20 ZHANG Bang-Hong, SHI Hai-Yan, WANG Ming-Hua. Chinese Journal of Pesticide Science, 2009, 11(2): 261~268
|
| [12] | 张帮红, 施海燕, 王鸣华. 农药学学报, 2009, 11(2): 261~268
|
| [13] | 3 Maienfisch P, Angst M, Brandl F, Fischer W, Hofer D, Kayser H, Kobel W, Rindlisbacher A, Senn R, Steinemann A, Widmer H. Pest Manag. Sci., 2001, 57(10): 906~913
|
| [14] | 4 ZHANG Guo-Sheng, HOU Guang-Xin. Pesticide Science and Administration, 2004, 25(3): 22~26
|
| [15] | 10 Singh S B, Mukherjee I, Maisnam J, Kumar P, Gopal M, Kulshrestha G. J. Agric. Food Chem., 2009, 57(23): 11277~11283
|
| [16] | 12 Kim H J, Liu S Z, Keum Y S, Li Q X. J. Agric. Food Chem., 2003, 51(7): 1823~1830
|
| [17] | 13 Ma H X, Xu Y J, Li Q X, Xu T, Wang X T, Li J. Food Addit. Contam., 2009, 26(5): 713~718
|
| [18] | 14 Xu T, Wei K Y, Wang J, Ma H X, Li J, Xu Y J, Li Q X. J. AOAC Int., 2010, 93(1): 12~18
|
| [19] | 15 Yu Y S, Sui H S, Han Z B, Li W, Luo M J, Tan J H. Cell Res., 2004, 14(4): 341~346
|
| [20] | 17 Wengatz I, Stoutamire D W, Gee S J, Hammock B D. J. Agric. Food Chem., 1998, 46(6): 2211~2221
|
| [21] | 18 Lee H J, Shan G, Ahn K C, Park E K, Watanabe T, Gee S J, Hammock B D. J. Agric. Food Chem., 2004, 52(5): 1039~1043
|
| [22] | 19 Shelver W L, Smith D J. J. Immunoassay, 2000, 21(1): 1~23
|
Full-Text
|
|
Contact Us
service@oalib.com QQ:3279437679 
WhatsApp +8615387084133
|
|
