The reagents used in the immunoassay may also cause matrix effects. Despite enormous advances in the design of immunoassays, unwanted interferences caused by matrix effects cannot be completely excluded. Moreover, the interferences in the measurement of different samples usually vary with each other, so when detecting a target pollutant of interest with an immunosensor, special methods for eliminating matrix effects must often be used to obtain correct assay results.Microcystin-LR (MC-LR) containing l-leucine and r-arginine in positions 2 and 4, respectively, is the most frequent and most toxic among nearly 80 microcystin variants obtained from Microcystis, Anabaena, Oscillatoria (Planktothrix), Nostoc and Anabaenopsis .
Many reported cases of animal-poisoning and human health diseases, some resulting in liver cancer and even death, are due to exposure to MCs via drinking and surface water [4�C6]. To minimize public exposure to MCs, the World Health Organization (WHO) has proposed a drinking water MC-LR guideline value (GV) of 1 ��g/L . Some immunoassay technologies have been developed to detect MC-LR [7,8], but due to the matrix interferences in water samples, most of them could not be applied to assay the real samples . Fluorescent immunosensors have been developed to determine various trace amounts of targets interest based on the principle of fluorescent immunoassay [10�C12]. However, a detailed evaluation of common organic and inorganic substances found in the environment for the detection of MC-LR based on fluorescent immunosensor is still missing.
We have previously introduced a new portable miniaturized evanescent wave all-fiber immunosensor (EWAI) to determine various trace amounts of targets interest based on the principle of immunoreaction and total internal reflect fluorescent (TIRF) . Here we use the slightly revised EWAI to investigate the influence of common interferences like PBS, pH, humic acid and copper ions on the sensitivity and stability of the MC-LR fluorescence immunoassay, and demonstrated that with the choice of a proper elimination method, the influence of interfering substances can be limited.2.?Experimental2.1. Immunoreagents and Chemicals3-mercaptopropyl-trimethoxysilane (MTS), ovalbumin (OVA), bovine serum albumin (BSA), N-(4-maleimidobutyryloxy) succinimide (GMBS), and 1-ethyl-3-(dimethylaminopropyl) carbodiimide hydrochloride (EDC) were purchased from Sigma-Aldrich (Steinheim, Germany).
MC-LR was obtained from Alexis (Lausen, Switzerland). Drug_discovery All the other reagents, unless specified, were supplied by Beijing Chemical Agents; these were also of analar grade and used without further purification. Distilled deionized water was used throughout the investigation. Monoclonal anti-MC-LR antibody (MC-LR-MAb. reference no.