Abstract
Fumonisin B1 (FMB1) is a food-born mycotoxin produced by Fusarium moniliforme. Monoclonal antibody against FMB1 (anti-FMB1 mAb) was produced in the hybridoma DV9, which was established from a BALB/c mouse immunized with bovine serum albumin conjugated FMB1 (FMB1-BSA). A competitive direct enzyme-linked immunosorbent assay (ELISA) showed that anti-FMB1 mAb has about 10 ppb of minimum FMB1 detection concentration and 220 ppb of 50% inhibition concentration (IC50). Much lower cross-reactivity of anti-FMB1 mAb on ochratoxin A, aflatoxin B1 and deoxynivalenol provided that anti-FMB1 mAb was specific for FMB1. The gene coding single chain variable fragment against FMB1 (anti-FMB1 scFv) was cloned from the hybridoma DV9 and was expressed in recombinant Escherichia coli. Insoluble anti-FMB1 scFv required optimization of its refolding condition, and hence functional scFv was obtained. By using indirect ELISA, about 12-fold lower binding activity of anti-FMB1 scFv on FMB1-BSA was obtained in comparison with that of the parental mAb.
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References
Abel S, Gelderblom WCA (1998) Oxidative damage and fumonisin B1-induced toxicity in primary rat hepatocytes and rat liver in vivo. Toxicology 131:121–131
Marasas WFO, Kellerman TS, Gelderblom WCA, Coetzer JAW, Thiel PG, Vanderlugt JJ (1988) Leukoencephalomalacia in a horse induced by fumonisin B1 isolated from Fusarium moniliforme. Onderstepoort J Vet Res 55:197–203
Colvin BM, Harrison LR (1992) Fumonisin-induced pulmonary edema and hydrothorax in swine. Mycopathologia 117:79–82
IARC (2002) IARC monographs on the evaluation of carcinogenic risks to humans. Some traditional herbal medicines, some mycotoxins, naphthalene and styrene. vol 82, IARC Press, Lyon
Castells M, Marin S, Sanchis V, Ramos AJ (2008) Distribution of fumonisins and aflatoxins in corn fractions during industrial cornflake processing. Int J Food Microbiol 123:81–87
Quan Y, Zhang Y, Wang S, Lee N, Kennedy IR (2006) A rapid and sensitive chemiluminescence enzyme-linked immunosorbent assay for the determination of fumonisin B1 in food samples. Anal Chim Acta 580:1–8
Chu FS, Lau HP, Fan TS, Zhang GS (1982) Ethylenediamine modified bovine serum albumin as protein carrier in the production of antibody against mycotoxins. J Immunol Methods 55:73–78
Hack R, Martlbauer E, Terplan G (1987) A monoclonal antibody to the trichothecene T-2 toxin: screening for the antibody by a direct enzyme immunoassay. J Vet Med Ser B-Infect Dis Vet Public Health 34:538–544
Shulman M, Wilde CD, Kohler G (1978) A better cell line for making hybridomas secreting specific antibodies. Nature 276:269–270
Cho YJ, Lee DH, Kim DO, Min WK, Bong KT, Lee GG, Seo JH (2005) Production of a monoclonal antibody against ochratoxin A and its application to immunochromatographic assay. J Agric Food Chem 53:8447–8451
Choi GH, Lee DH, Min WK, Cho YJ, Kweon DH, Son DH, Park K, Seo JH (2004) Cloning, expression, and characterization of single-chain variable fragment antibody against mycotoxin deoxynivalenol in recombinant Escherichia coli. Protein Expr Purif 35:84–92
Son YJ, Park KH, Lee SY, Oh SJ, Kim CK, Choi PT, Park YC, Seo JH (2007) Effects of temperature shift strategies on human preproinsulin production in the fed-batch fermentation of recombinant Escherichia coli. Biotechnol Bioprocess Eng 5:556–561
Haaijman JJ, Coolen J, Krose CJM, Pronk GJ, Ming ZF (1986) Fluorescein and tetramethyl rhodamine as haptens in enzyme immunohistochemistry. Histochem Cell Biol 84:363–370
Candlish AAG, Smith JE, Stimson WH (1989) Monoclonal antibody technology for mycotoxins. Biotechnol Adv 7:401–418
Yang X, Hu W, Li F, Xia H, Zhang Z (2005) Gene cloning, bacterial expression, in vitro refolding, and characterization of a single-chain Fv antibody against PreS1(21–47) fragment of HBsAg. Protein Expr Purif 41:341–348
Kipriyanov SM, Moldenhauer G, Martin AC, Kupriyanova OA, Little M (1997) Two amino acid mutations in an anti-human CD3 single chain Fv antibody fragment that affect the yield on bacterial secretion but not the affinity. Protein Eng Des Sel 10:445–453
Guo JQ, Li QM, Zhou JY, Zhang GP, Yang YY, Xing GX, Zhao D, You SY, Zhang CY (2006) Efficient recovery of the functional IP10-scFv fusion protein from inclusion bodies with an on-column refolding system. Protein Expr Purif 45:168–174
Harrison JS, Keshavarz-Moore E (1996) Production of antibody fragments in Escherichia coli. Ann NY Acad Sci 782:143–158
Chiarappa P, Baudrimont I, Borracci P, Carratu MR, Creppy EE (2004) Evaluation of possible synergistic effects of ochratoxin A and fumonisin B1 in CaCo2, C6 and VERO cells. Toxicol Appl Pharmacol 197:312–313
Han MJ, Seo YK, Yoon HH, Song KY, Park JK (2008) Effect of mechanical tension on the human dental pulp cells. Biotechnol Bioprocess Eng 13:410–417
Joung HA, Shim WB, Chung DW, Ahn JH, Chung BH, Choi HS, Ha SD, Kim KS, Lee KH, Kim CH, Kim KY, Kim MG (2007) Screening of a specific monoclonal antibody against and detection of Listeria monocytogenes whole cells using a surface plasma resonance biosensor. Biotechnol Bioprocess Eng 12:80–85
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This study was supported by a grant (Code number 20070301034013) from BioGreen 21 Program, Rural Development Administration, and WCU (World Class University) program through the Korea Science and Engineering Foundation (Grant No. 08-0238).
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Min, WK., Cho, YJ., Park, JB. et al. Production and characterization of monoclonal antibody and its recombinant single chain variable fragment specific for a food-born mycotoxin, fumonisin B1 . Bioprocess Biosyst Eng 33, 109–115 (2010). https://doi.org/10.1007/s00449-009-0350-9
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DOI: https://doi.org/10.1007/s00449-009-0350-9