Production and characterization of a monoclonal antibody against domoic acid and its application to enzyme immunoassay

Toxicon. 1999 Nov;37(11):1579-89. doi: 10.1016/s0041-0101(99)00106-3.

Abstract

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.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / biosynthesis*
  • Antibodies, Monoclonal / chemistry*
  • Antibody Specificity
  • Bivalvia / chemistry
  • Brachyura / chemistry
  • Cattle
  • Cross Reactions
  • Foodborne Diseases
  • Humans
  • Immunoenzyme Techniques
  • Isomerism
  • Kainic Acid / analogs & derivatives*
  • Kainic Acid / analysis
  • Kainic Acid / immunology
  • Marine Toxins / analysis*
  • Marine Toxins / immunology*
  • Mice
  • Mice, Inbred BALB C
  • Ostreidae / chemistry
  • Ovalbumin / chemistry
  • Serum Albumin, Bovine / chemistry
  • Shellfish / analysis
  • gamma-Globulins / chemistry

Substances

  • Antibodies, Monoclonal
  • Marine Toxins
  • gamma-Globulins
  • Serum Albumin, Bovine
  • Ovalbumin
  • domoic acid
  • Kainic Acid