Production and specificity of mono and polyclonal antibodies against microcystins conjugated through N-methyldehydroalanine.
Microcystins (MCs) are a group of closely related toxic cyclic heptapeptides produced by common cyanobacteria (blue-green algae). Their toxicity is associated with specific inhibition of intracellular protein phosphatases type-1 and type-2A (PP1 and PP2A, respectively). We have developed a battery of antibodies to microcystins using chemical modification (aminoethylation) of one of its core amino acids, N-methyl-dehydroalanine. The developed antibodies displayed different reactivities to closely related MCs. Selected monoclonal antibodies were used for quantitative competitive ELISA assays. The analytical sensitivity of these assays was up to 1 ng/ml. Comparison of the developed ELISA tests with HPLC-based measurements of MCs in laboratory and field samples showed a good correspondence between the results yielded by these two methods. The antibodies developed by this technique provide the means for developing extremely sensitive and specific analytical assays for direct measurement of toxins in cyanobacterial or water samples.
Mikhailov
A
A
Department of Biology, Laboratory of Animal Physiology, University of Turku, FIN-20014, Turku, Finland.
Härmälä-Braskén
A S
AS
Meriluoto
J
J
Sorokina
Y
Y
Dietrich
D
D
Eriksson
J E
JE
D000409:Alanine / Q000031:analogs & derivatives; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D001427:Bacterial Toxins / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D052998:Microcystins; D010456:Peptides, Cyclic / Q000032:analysis / Q000276:immunology*; D011817:Rabbits
D000906:Antibodies; D000911:Antibodies, Monoclonal; D001427:Bacterial Toxins; D052998:Microcystins; D010456:Peptides, Cyclic; C078588:microcystin; C037064:N-methyldehydroalanine; D000409:Alanine