Production and characterization of a monoclonal antibody against domoic acid and its application to enzyme immunoassay.
For production of monoclonal antibodies against domoic acid, a causative agent of amnesic shellfish poisoning, three immunogens, domoic acid conjugated with bovine serum albumin (BSA), ovalbumin (OVA) and human gamma globulin (HGG), were prepared. The antiserum obtained from BALB/c mice immunized with domoic acid-BSA showed the highest affinity for domoic acid. The monoclonal antibody, DA-3, obtained from the mice was highly specific for domoic acid and showed a minor cross-reactivity with the isomers of domoic acid (isodomoic acids B, E, F, G and H), except for isodomoic acid A. Using DA-3 antibody, an indirect competitive enzyme immunoassay (idc-EIA) was developed for measurement of domoic acid. The working range for quantitative measurement of domoic acid and the quantification limit for domoic acid in shellfish were estimated to be 0.15-10 ng/ml and less than 0.04 microg/g, respectively. The mean recovery of domoic acid added to extracts of shellfish at toxin levels of 0.02 to 0.2 microg/ml was 103% with a coefficient of variation of 4.5%. The newly developed idc-EIA seems to be a useful method for monitoring domoic acid in shellfish.
Kawatsu
K
K
Department of Food Microbiology, Osaka Prefectural Institute of Public Health, Japan. kawatu@iph.pref.osaka.jp
Hamano
Y
Y
Noguchi
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry*; D000918:Antibody Specificity; D049872:Bivalvia / Q000737:chemistry; D003386:Brachyura / Q000737:chemistry; D002417:Cattle; D003429:Cross Reactions; D005517:Foodborne Diseases; D006801:Humans; D007124:Immunoenzyme Techniques; D007536:Isomerism; D007608:Kainic Acid / Q000031:analogs & derivatives* / Q000032:analysis / Q000276:immunology; D008387:Marine Toxins / Q000032:analysis* / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D010125:Ostreidae / Q000737:chemistry; D010047:Ovalbumin / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry; D012758:Shellfish / Q000032:analysis; D005719:gamma-Globulins / Q000737:chemistry
D000911:Antibodies, Monoclonal; D008387:Marine Toxins; D005719:gamma-Globulins; D012710:Serum Albumin, Bovine; D010047:Ovalbumin; C012301:domoic acid; D007608:Kainic Acid