Production and characterization of monoclonal antibody and its recombinant single chain variable fragment specific for a food-born mycotoxin, fumonisin B1.
Fumonisin B(1) (FMB(1)) is a food-born mycotoxin produced by Fusarium moniliforme. Monoclonal antibody against FMB(1) (anti-FMB(1) mAb) was produced in the hybridoma DV9, which was established from a BALB/c mouse immunized with bovine serum albumin conjugated FMB(1) (FMB(1)-BSA). A competitive direct enzyme-linked immunosorbent assay (ELISA) showed that anti-FMB(1) mAb has about 10 ppb of minimum FMB(1) detection concentration and 220 ppb of 50% inhibition concentration (IC(50)). Much lower cross-reactivity of anti-FMB(1) mAb on ochratoxin A, aflatoxin B(1) and deoxynivalenol provided that anti-FMB(1) mAb was specific for FMB(1). The gene coding single chain variable fragment against FMB(1) (anti-FMB(1) scFv) was cloned from the hybridoma DV9 and was expressed in recombinant Escherichia coli. Insoluble anti-FMB(1) 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-FMB(1) scFv on FMB(1)-BSA was obtained in comparison with that of the parental mAb.
Min
Won-Ki
WK
Department of Agricultural Biotechnology, Seoul National University, Seoul 151-921, Korea.
Cho
Young-Jin
YJ
Park
Jun-Bock
JB
Bae
Yi-Hyun
YH
Kim
Eun-Jeong
EJ
Park
Kyungmoon
K
Park
Yong-Cheol
YC
Seo
Jin-Ho
JH
D016604:Aflatoxin B1 / Q000737:chemistry / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D000915:Antibody Affinity / Q000235:genetics / Q000276:immunology; D000918:Antibody Specificity / Q000235:genetics / Q000276:immunology*; D002417:Cattle; D003429:Cross Reactions / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D037341:Fumonisins / Q000737:chemistry / Q000276:immunology*; D005670:Fusarium / Q000737:chemistry; D015870:Gene Expression; D051379:Mice; D008807:Mice, Inbred BALB C; D009793:Ochratoxins / Q000737:chemistry / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000276:immunology; D057127:Single-Chain Antibodies / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D014255:Trichothecenes / Q000737:chemistry / Q000276:immunology
D000911:Antibodies, Monoclonal; D037341:Fumonisins; D009793:Ochratoxins; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D014255:Trichothecenes; C025589:ochratoxin A; D012710:Serum Albumin, Bovine; C056933:fumonisin B1; D016604:Aflatoxin B1; C007262:deoxynivalenol