Monoclonal antibody specific for cyanoginosin-LA: preparation and characterization

The toxin cyanoginosin-LA (MW 909), isolated from Microcystis aeruginosa, was successfully conjugated with polylysine and muramyl dipeptide to form a high molecular weight complex consisting of a hapten, a carrier and a built-in adjuvant. This complex was used for the immunization of mice. Monoclonal antibodies specific for cyanoginosin-LA were produced using the hybridoma technique. The ten most efficient producers of these antibodies were further characterized and the monoclonal antibody produced by them was found to be identical on the basis of additivity test, isoelectric focussing and sub-class immunoglobulin typing results. The anti-cyanoginosin-LA monoclonal antibody focussed at a pH of approximately 6.55 and was found to belong to the mouse immunoglobulin subclass IgM. Large scale production of the antibody (in vivo) was followed by purification with ammonium sulphate precipitation and high performance liquid chromatography. The anticyanoginosin-LA monoclonal antibody, when reacted with six variants of cyanoginosin (other than cyanoginosin-LA), bound all with equal efficiency.