Determination of trace amount of cyanobacterial toxin in water by microchip based enzyme-linked immunosorbent assay.
Routine monitoring of microcystin in natural waters is difficult because the concentration of the toxin is usually lower than the detection limits. As a more sensitive detection method for microcystin, we developed a microchip based enzyme-linked immunosorbent assay (ELISA) based on monoclonal antibodies. New monoclonal antibodies against the microcystin leucine-arginine variant (MCLR), a cyclic peptide toxin of the freshwater cyanobacterium Microcystis aeruginosa, were prepared from cloned hybridoma cell lines. We used keyhole limpet hemocyanin(KLH)-conjugated MCLR as an immunogen for the production of mouse monoclonal antibody. The immunization, cell fusion, and screening of hybridoma cells producing anti-MCLR antibody were conducted. Since the ELISA test was highly sensitive, the newly developed microchip based ELISA can be suitable for the trace analysis of cyanobacterial hepatotoxins, microcystins in water. The linear responses of monoclonal antibodies with different concentrations of microcystin LR were established between 0.025 and 0.3 ng/mL.
Pyo
Dongjin
D
Department of Chemistry, Kangwon National University, ChunChon, South Korea. pyod@kangwon.ac.kr
Hahn
Jong Hoon
JH
D000911:Antibodies, Monoclonal; D001427:Bacterial Toxins / Q000032:analysis; D000087522:Cyanobacteria Toxins; D004784:Environmental Monitoring / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D008387:Marine Toxins / Q000032:analysis; D046208:Microchip Analytical Procedures; D052998:Microcystins / Q000032:analysis*; D046931:Microcystis / Q000276:immunology / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D014867:Water / Q000032:analysis*; D014871:Water Microbiology
D000911:Antibodies, Monoclonal; D001427:Bacterial Toxins; D000087522:Cyanobacteria Toxins; D008387:Marine Toxins; D052998:Microcystins; D014867:Water; C078588:microcystin