title: Production of antibodies against microcystin-RR for the assessment of purified microcystins and cyanobacterial environmental samples.
abstract: Microcystins (MC) are cyanobacterial hepatotoxins responsible for animal-poisoning and human health incidents. Immunoassays provide a sensitive means to detect these toxins, although cross-reactivity characteristics of different antibodies are variable, and most antibodies have been produced against MC-LR. Here, we have produced the first polyclonal antibodies against the commonly occurring variant, MC-RR, and compared them with MC-LR antibodies for the analysis of purified MCs and cyanobacterial environmental samples. Both antisera cross-reacted with all MCs tested, and with the related cyanobacterial hepatotoxin nodularin-R, but not with non-toxic cyanobacterial peptides. In general, better cross-reactivity characteristics were observed with the MC-RR antisera and limits of quantification were lower for most variants, with all MCs tested and nodularin-R having limits of quantification of 0.31 nM or below. The antisera had different affinities to mixtures containing pooled MC-LR and MC-RR, with MC-LR antisera underestimating total MC concentration when MC-RR represented over 70% of the total MC pool. Both antisera correlated well with HPLC-UV data when incorporated into ELISAs to screen previously characterised environmental samples from Aland, Finland. MC-RR antisera are useful for screening samples containing multiple MCs, and particularly for samples primarily containing MC-RR variants.
authors.affiliation: Division of Environmental and Applied Biology, School of Life Sciences, University of Dundee, Dundee DD1 4HN, UK. f.m.young@dundee.ac.uk <f.m.young@dundee.ac.uk>
authors.forename: Fiona M
authors.initials: FM
authors.lastname: Young
authors.forename: James S
authors.initials: JS
authors.lastname: Metcalf
authors.forename: Jussi A O
authors.initials: JA
authors.lastname: Meriluoto
authors.forename: Lisa
authors.initials: L
authors.lastname: Spoof
authors.forename: Louise F
authors.initials: LF
authors.lastname: Morrison
authors.forename: Geoffrey A
authors.initials: GA
authors.lastname: Codd
mesh_terms: D000907:Antibodies, Bacterial / Q000096:biosynthesis*; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D000458:Cyanobacteria / Q000276:immunology / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay; D007106:Immune Sera; D008387:Marine Toxins; D052998:Microcystins; D010456:Peptides, Cyclic / Q000276:immunology*; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet
chemical_list: D000907:Antibodies, Bacterial; D007106:Immune Sera; D008387:Marine Toxins; D052998:Microcystins; D010456:Peptides, Cyclic; C063855:microcystin RR