伏马菌素B1单克隆抗体的制备及鉴定

doi:10.3864/j.issn.0578-1752.2011.20.019

Abstract

Abstract: 【Objective】 Mycotoxins can lead to serious food security issues. This study focused on the generation of the specific monoclonal antibody (McAb) against fumonisin B1. 【Method】 The artificial antigen of fumonisin B1 was prepared and a hybridoma cell line was screened with method of hybridoma. Monoclonal antibody was generated by murine ascites and identified. 【Result】 A hybridoma cell line of F3 was screened. The antibody secreted by F3 was classified into IgG1 and the light chain type was κ; the results of 10% SDS-PAGE showed that the molecular weight of high chain was 50 kD and the light chain was 25 kD, respectively. The titer of McAb was 1:3 200 in supernatant of F3 and 1:51 200 in ascites with indirect enzyme-linked immunosorbent assay. The affinity was 1.60×10-8 mol•L-1 and the value of IC50 was 3.589 ng•mL-1. There were no cross reactions to other mycotoxins. The high specificity of antibody was identified by western blotting. 【Conclusion】The McAb against FB1 in this experiment are of sensitivity and specificity.

Key words: fumonisinB1, artificialantigen, monoclonalantibody, enzyme-linkedimmunosorbentassay

WANG Ying, ZHENG Hao, HE Cheng-Hua, ZHANG Ai-Hua, FENG Lu-Lu, HUANG Chao-Ying, ZHANG Hai-Bin. Generation and Identification of the Monoclonal Antibody Against Fumonisin B1[J].Scientia Agricultura Sinica, 2011, 44(20): 4302-4308.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.chinaagrisci.com/EN/10.3864/j.issn.0578-1752.2011.20.019

https://www.chinaagrisci.com/EN/Y2011/V44/I20/4302

References

[1]Sydenham E W, Gelderblom W C A, Thiel P G, Marasas W F O. Evidence for the natural occurrence of fumonisin B1, a mycotoxin produced by Fusarium moniliforme, in corn. Journal of Agricultural and Food Chemistry, 1990, 38: 285-290.

[2]刘书宇, 杨美华. 伏马菌素的研究进展. 安徽农业科学, 2009, 37(24): 11397-11399.

Liu S Y, Yang M H. Research progress on Fumonisins. Journal of Anhui Agricultural Science, 2009, 37(24): 11397-11399. (in Chinese)

[3]Shephard G S, Thiel P G, Stockenstrom S, Sydenham E W. Worldwide survey of fumonisin contamination of corn and corn-based products. Journal of AOAC International, 1996, 79(3): 671-687.

[4]宫慧芝, 那磊, 李岩溪, 计融, 杨军. 玉米中伏马菌素B1高效液相色谱检测. 中国公共卫生, 2008, 24(3): 381-382.

Gong H Z, Na L, Li Y X, Ji R, Yang J. Determination of fumonisins B1 in corns by high performance liquid chromatography. Chinese Journal Public Health, 2008, 24(3): 381-382. (in Chinese)

[5]Shephard G S, Sydenham E W, Thiel P G, Gelderblom W C A. Quantitative determination of fumonisins B1 and B2, by high performance liquid chromatography with fluorescence detection. Journal of Liquid Chromatography, 1990, 13: 2077-2087.

[6]Shelby R A, Rottinghaus G E, Minor H C. Comparison of thin-layer chromatography and competitive immunoassay methods for detecting fumonisin on maize. Journal of Agricultural and Food Chemistry, 1994, 42: 2064-2067.

[7]Maragos C M. Capillary zone electrophoresis and HPLC for the analysis of fluoresce in isothiocyanate-labelled fumonisin B1. Journal of Agricultural and Food Chemistry, 1995, 43: 390-394.

[8]Royer D, Humpf H U, Guy P A. Quantitative analysis of Fusarium mycotoxins in maize using accelerated solvent extraction before liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry. Food Additives and Contaminants, 2004, 21(7): 678-692.

[9]薛秋艳, 徐剑宏, 祭芳, 史建荣. 玉米中伏马毒素FB1液相色谱电喷雾串联质谱测定方法的建立. 江苏农业学报, 2010, 26(3): 627-630.

Xue Q Y, Xu J H, Ji F, Shi J R. Establishment of high performance liquid chromatography-electrospray ionization tandem mass spectrometry for detecting fumonisin B1 in maize. Jiangsu Journal of Agricultural Science, 2010, 26(3): 627-630. (in Chinese)

[10]Plattner R D, Norred W P, Bacon C W, Voss K A, Peterson R, Shackelford D D, Weisleder D. A method of detection of fumonisins in corn samples associated with field cases of equine leukoencephalomalacia. Mycologia, 1990, 82: 698-702.

[11]刘师文, 何庆华, 邹龙, 许杨. 谷物中伏马菌素B1酶联免疫分析法的建立. 食品科学, 2010, 31(18): 350-354.

Liu S W, He Q H, Zou L, Xu Y. Development of an ELISA method for determining fumonisin B1 in cereals. Food Science, 2010, 31(18): 350-354. (in Chinese)

[12]Yeung J M, Perlusky D B, Savard M E, Dang B D M, Robinson L A. Sensitive immunoassay for fumonisin B1 in corn. Journal of Agricultural and Food Chemistry, 1996, 44: 3582-3586.

[13]Christensen H R, Yu F Y, Chu F S. Development of a polyclonal antibody-based sensitive enzyme linked immunosorbent assay for fumonisin B4. Journal of Agricultural and Food Chemistry, 2000, 48: 1977-1984.

[14]林金明, 赵利霞. 化学发光免疫分析. 北京: 化学工业出版社, 2008: 109.

Lin J M, Zhao L X. Chemiluminescent Immunoassay. Beijing: Chemical Industry Press, 2008: 109. (in Chinese)

[15]Quan Y, Zhang Y, Wang S, Lee N J, Kennedy I R. A rapid and sensitive chemiluminescence enzyme-linked immunosorbent assay for the determination of fumonisin B1 in food samples. Analytica Chimica Acta, 2006, (580): 1-8.

[16]刘秀梵. 单克隆抗体在农业上的应用. 合肥: 安徽科学技术出版社, 1994: 18.

Liu X F. Monoclonal Antibody Applications in Agriculture. Hefei: Anhui Science and Technology Press, 1994: 18. (in Chinese)

[17]冯仁青, 郭振泉, 宓捷波. 现代抗体技术及其应用. 北京: 北京大学出版社, 2006: 198.

Feng R Q, Guo Z Q, Mi J B. Modern Antibody Technology and Its Applications. Beijing: Beijing University Press, 2006: 198. (in Chinese)

[18]万文徽. 单克隆抗体亲和常数的测定. 单克隆抗体通讯, 1993, 9(2): 72-75.

Wan W H. Determination of the affinity constant for monoclonal antibody. Journal of Monoconal Antibody, 1993, 9(2): 72-75. (in Chinese)

[19]Zhu G N, Gui W J, Zheng Z T, Cheng J L, Wu G. Synthesis and identification of artificial antigen for imidacloprid. Agricultural Sciences in China, 2006, 5(4): 307-312.

[20]张梅, 张继瑜, 李剑勇. 兽药人工抗原合成的研究进展. 中兽医医药杂志, 2006, 1: 63-65.

Zhang M, Zhang J Y, Li J Y. Research progress on the synthesis of artificial antigen on animal remedy. Journal of Traditional Chinese Veterinary Medicine, 2006, 1: 63-65. (in Chinese)

[21]祭芳, 陈正贤, 徐剑宏, 陆琼娴, 史建荣. 抗脱氧雪腐镰刀菌烯醇单克隆抗体的制备. 微生物学报, 48(7): 929-934.

Ji F, Chen Z X, Xu J H, Lu Q X, Shi J R. Development of the monoclonal antibody to deoxynivalenol. Acta Microbiologica Sinica, 48(7): 929-934. (in Chinese)

[22]杨金易, 吴青, 王弘, 潘科, 雷红涛, 肖治理, 郑晓锋, 孙远明. 高亲和力的农药克百威单克隆抗体的制备及鉴定. 中国农业科学, 2007, 40(3): 518-523.

Yang J Y, Wu Q, Wang H, Pan K, Lei H T, Xiao Z L, Zheng X F, Sun Y M. Production and identification of high affinity monoclonal antibodies against pesticide carbofuran. Scientia Agricultura Sinica, 2007, 40(3): 518-523. (in Chinese)

[23]职爱民, 李青梅, 刘庆堂, 刘宣兵, 杨继飞, 杨苏珍, 柴书军, 杨艳艳, 邓瑞广, 张改平. 抗庆大霉素单克隆抗体的制备及其初步应用. 中国农业科学, 2010, 43(12): 2584-2589.

Zhi A M, Li Q M, Liu Q T, Liu X B, Yang J F, Yang S Z, Chai S J, Yang Y Y, Deng R G, Zhang G P. Preparation and preliminary application of anti-gentamicin monoclonal antibody. Scientia Agricultura Sinica, 2010, 43(12): 2584-2589. (in Chinese)

[24]胡鲲, 黄宣运, 姜有声, 方伟, 杨先乐. 环丙沙星单克隆抗体的制备及其免疫学特性分析. 中国免疫学杂志, 2010, 26(6): 538-543.

Hu K, Huang X Y, Jiang Y S, Fang W, Yang X L. Generation of the monoclonal antibody against ciprofloxacin and analysis for its immunological traits. Chinese Journal of Immunology, 2010, 26(6): 538-543. (in Chinese)

[25]姜金庆, 张海棠, 王自良, 王建华, 范国英. 19-去甲睾酮异源性ciELISA试剂盒的研制及应用. 中国生物工程杂志, 2010, 30(9): 68-74.

Jiang J Q, Zhang H T, Wang Z L, Wang J H, Fan G Y. Development and application of heterologous ciELISA kit for rapid detection of 19-nortestosterone residues. China Biotechnology, 2010, 30(9): 68-74. (in Chinese)

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

Comments

Recommended 0

No Suggested Reading articles found!

Generation and Identification of the Monoclonal Antibody Against Fumonisin B1

PDF

Cited

Abstract

Cite this article

share this article

References

Related Articles 0

Metrics

Comments

Recommended 0