A highly specific immunoassay for microcystin-LR detection based on a monoclonal antibody.
Microcystins (MC) are cyanobacterial hepatotoxins responsible for animal-poisoning and human health incidents. Immunoassays provide a sensitive and fast means to detect these toxins, but cross-reactivity (CR) characteristic of different antibodies was variable. Here, we have produced and characterized a monoclonal antibody (Clone MC8C10) with highly specificity against the most frequent and most toxic variant of microcystins, MC-LR. MC8C10 is more specific against MC-LR among the reported antibodies before. The immunogen was synthesized from the modified MC-LR and bovine serum albumin (BSA). An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) with MC8C10 was established to detect the MCs in waters, which showed highly specificity with MC-LR, and have a detection limit for MC-LR 0.1 microg L(-1), the 50% inhibition concentration (IC50) for MC-LR was 1.8+/-0.1 microg L(-1) and the quantitative detection range was from 0.3 to 10 microg L(-1). The [4-arginine] microcystins and the nodularin-R showed lower cross-reactivates (CR<10%), and other MCs such as MC-LF and MC-LW are not recognized (CR<10(-4)). The analysis results of real water samples with ic-ELISA showed that all the coefficients of variation were less than 15%, and the recovery was (100.3+/-5.9)%. So the highly specific ic-ELISA will commendably suit for sensitive analysis for MC-LR in surface water as well as drinking water.
Sheng
Jian-Wu
JW
Environmental Simulation and Pollution Control State Key Joint Laboratory, Department of Environmental Science and Engineering, Tsinghua University, Beijing 100084, China.
He
Miao
M
Shi
Han-Chang
HC
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006825:Hybridomas; D008387:Marine Toxins; D051379:Mice; D008807:Mice, Inbred BALB C; D052998:Microcystins / Q000032:analysis* / Q000276:immunology; D010456:Peptides, Cyclic / Q000032:analysis* / Q000276:immunology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013154:Spleen / Q000166:cytology / Q000276:immunology; D014873:Water Pollutants / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D008387:Marine Toxins; D052998:Microcystins; D010456:Peptides, Cyclic; D014873:Water Pollutants; C063998:nodularin; C057862:cyanoginosin LR