Keywords
How to Cite
Abstract
Objective:To acquire monoclonal hybridoma cell lines against fummonisin B1(FB1) and extract monoclonal antibody against FB1.
Methods: Coupling antigens of FB1-KLH and FB1-BSA with chemical methods and immune 6-8 weeks old femaleBALB/c mice with FB1-KLH. Integrating spleen cells with sp2/0 myeloma cells to acquire hybridoma cell lines secreting McAb against FB1. The method of multiple subclones was used to select cell lines stably secreting McAb. McAbs was got from ascites and purified. The subclass of antibody was measured and the molecular weight was identified. The specificity and sensitivity of McAb were identified with indirect competitive inhibition ELISA.
Results:The results of serum from immuned mice showed that after five times of immunization the titer stables at 1×10-6, and the McAb belongs to IgG1 subclass, the light chain was ?, the molecular weight of heavy and light chain were 55kDa and 32kDa, respectively. ELISA results showed that McAb could react with FB1. The linear range indirect competitive inhibition ELISA is 10-500ng/ml.
Conclusion:The monoclonal hybridoma cell lines and the high specificity,high sensitivity of FB1-McAb was got.
References
Guo Y, Liu X, Liu J. Studies on immunoassay for detecting fumonisin B1. Wei Sheng Yan Jiu 1999; 28(4): 238-240.
Mansur AR, Yu CC, Oh DH. Efficiency of gamma irradiation to inactivate growth and fumonisin production of Fusarium moniliforme on corn grains. J Microbiol Biotechnol 2014; 24(2): 209-216. http://dx.doi.org/10.4014/jmb.1307.07079
Yoshizawa T, Yamashita A, Luo Y. Fumonisin occurrence in corn from high- and low-risk areas for human esophageal cancer in China. Appl Environ Microbiol 1994; 60(5): 1626-1629.
Huang C, Dickman M, Henderson G, Jones C. Repression of protein kinase C and stimulation of cyclic AMP response elements by fumonisin, a fungal encoded toxin which is a carcinogen. Cancer Res 1995; 55(8): 1655-1659.
Ramljak D, Calvert RJ, Wiesenfeld PW, Diwan BA, Catipovic B, Marasas WF, Victor TC, Anderson LM, Gelderblom WC. A potential mechanism for fumonisin B(1)-mediated hepatocarcinogenesis: cyclin D1 stabilization associated with activation of Akt and inhibition of GSK-3beta activity. Carcinogenesis 2000; 21(8): 1537-1546. http://dx.doi.org/10.1093/carcin/21.8.1537
Turner PC, Nikiema P, Wild CP. Fumonisin contamination of food: progress in development of biomarkers to better assess human health risks. Mutat Res 1999; 443(1-2): 81-93. http://dx.doi.org/10.1016/S1383-5742(99)00012-5
De Girolamo A, Lattanzio VM, Schena R, Visconti A, Pascale M. Use of liquid chromatography-high-resolution mass spectrometry for isolation and characterization of hydrolyzed fumonisins and relevant analysis in maize-based products. J Mass Spectrom 2014; 49(4): 297-305. http://dx.doi.org/10.1002/jms.3342
Wild CP, Gong YY. Mycotoxins and human disease: a largely ignored global health issue. Carcinogenesis 2010; 31(1): 71-82. http://dx.doi.org/10.1093/carcin/bgp264
Marin S, Ramos AJ, Cano-Sancho G, Sanchis V. Mycotoxins: occurrence, toxicology, and exposure assessment. Food Chem Toxicol 2013; 60: 218-237. http://dx.doi.org/10.1016/j.fct.2013.07.047
Sharma N, He Q, Sharma RP. Amelioration of fumonisin B1 hepatotoxicity in mice by depletion of T cells with anti-Thy-1.2. Toxicology 2006; 223(3): 191-201. http://dx.doi.org/10.1016/j.tox.2006.03.021
Ren ZQ, Liu TC, Zhuang SH, Lin GF, Hou JY, Wu YS. Establishment of Magnetic Microparticles-Assisted Time-Resolved Fluoroimmunoassay for Determinating Biomarker Models in Human Serum. PLoS One 2015; 10(6): e0130481. http://dx.doi.org/10.1371/journal.pone.0130481
Lamberti I, Tanzarella C, Solinas I, Padula C, Mosiello L. An antibody-based microarray assay for the simultaneous detection of aflatoxin B1 and fumonisin B1. Mycotoxin Res 2009; 25(4): 193-200. http://dx.doi.org/10.1007/s12550-009-0028-9
Darwish IA, Alzoman NZ, Abuhejail RM, El-Samani TE. Synthesis of hapten and preparation of specific polyclonal antibody with high affinity for lenalidomide, the potent drug for treatment of multiple myeloma. Chem Cent J 2012; 6(1): 125. http://dx.doi.org/10.1186/1752-153X-6-125
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Copyright (c) 2015 Guangyu Yang, Wenxue Li, Xiaoyun Liang, Jinyin Wu , Wei Zhu