Generation of antibodies directed against the low-immunogenic peptide-toxins microcystin-LR/RR and nodularin

Int J Immunopharmacol. 2000 May;22(5):339-53. doi: 10.1016/s0192-0561(99)00086-7.

Abstract

The preparation of antibodies against the liver toxin microcystin, as described here, is of major importance for its detection and purification in food and water, and for a therapeutic approach to neutralize the toxin by passive immunization. Microcystin-LR (MLR) and microcystin-RR (MRR) were purified from cyanobacterial cell materials by extraction, Sephadex LH-20-, ODS silica gel-, ionic exchange and RP-HPLC-chromatography. In order to reduce the toxicity for parenteral administration, microcystins were coupled by the carbodiimide method to poly-L-lysine (PLL(50.000)). Mice and rabbits were immunized with the conjugates in the presence of two lipopeptide immunoadjuvants (P(3)CSK(4) and P(3)CS-T(h)). High MLR-specific antibody levels were observed after parenteral coadministration of antigen and lipopeptides, whereas no anti-MLR antibodies were obtained with free microcystin or the microcystin-PLL(50.000)-conjugate in the absence of lipopeptide. In oral immunization, coadministration of antigen and adjuvants resulted in an accelerated development of anti MLR-specific antibodies and high antibody levels. Using the antisera, we could detect different microcystins and nodularin down to a concentration range of 10-50 ng/ml by a competitive inhibition ELISA; detection of microcystins in crude cell preparations was also possible. Furthermore, microcystins from different sources could be detected and discriminated from cyclic cyanopeptolines.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Administration, Oral
  • Animals
  • Antibody Formation
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Immune Sera / immunology
  • Immunization
  • Marine Toxins
  • Mice
  • Mice, Inbred BALB C
  • Microcystins
  • Peptides, Cyclic / immunology*
  • Peptides, Cyclic / isolation & purification
  • Rabbits

Substances

  • Immune Sera
  • Marine Toxins
  • Microcystins
  • Peptides, Cyclic
  • nodularin
  • microcystin
  • cyanoginosin LR