Generation of antibodies reactive with fumonisins B1, B2, and B3 by using cholera toxin as the carrier-adjuvant.
Murine polyclonal antibodies reactive with fumonisins B1, B2, and B3 were produced after a novel immunization procedure with cholera toxin as both a hapten carrier protein and adjuvant. Immunization of mice with two 7.5-micrograms doses of fumonisin B1-cholera toxin conjugate without adjuvant resulted in the production of fumonisin B1-specific antibodies in all mice within 15 days when intraperitoneal, subcutaneous, and intravenous routes were used. In contrast, conventional immunization procedures with fumonisin B1-bovine serum albumin conjugates with and without Freund's adjuvant were largely ineffective. Fumonisin antibodies could be readily mass-produced in ascites fluid by using cholera toxin as a carrier-adjuvant. A competitive indirect enzyme-linked immunosorbent assay (ELISA) was devised whereby immobilized fumonisin B1-ovalbumin and free fumonisin B1 competed for antibody binding. The detection limit for fumonisin B1 in the ELISA was 100 ng/ml. The antiserum cross-reacted with fumonisins B2 and B3 but not with the hydrolyzed backbone of fumonisin B1 and tricarballylic acid. Concentrations of fumonisins B1, B2, and B3 required for 50% binding inhibition were 260, 300, and 650 ng/ml, respectively. These polyclonal antibodies should find wide usage in the ELISA for fumonisins in foods, feeds, and tissues.
Azcona-Olivera
J I
JI
Department of Food Science and Human Nutrition, Michigan State University, East Lansing 48824-1224.
Abouzied
M M
MM
Plattner
R D
RD
Norred
W P
WP
Pestka
J J
JJ
D000276:Adjuvants, Immunologic; D000906:Antibodies / Q000276:immunology; D002772:Cholera Toxin / Q000276:immunology; D004337:Drug Carriers; D004797:Enzyme-Linked Immunosorbent Assay; D037341:Fumonisins; D009183:Mycotoxins / Q000276:immunology*
D000276:Adjuvants, Immunologic; D000906:Antibodies; D004337:Drug Carriers; D037341:Fumonisins; D009183:Mycotoxins; C056934:fumonisin B2; C073063:fumonisin B3; C056933:fumonisin B1; D002772:Cholera Toxin