This work reports a novel electrochemical immunoassay protocol with signal amplification for determination of proteins (human IgG here used as a model target analyte) at an ultralow concentration using DNA-based hybridization chain reaction (HCR). The immuno-HCR assay consists of magnetic immunosensing probes, nanogold-labeled signal probes conjugated with the DNA initiator strands, and two different hairpin DNA molecules. The signal is amplified by the labeled ferrocene on the hairpin probes. In the presence of target IgG, the sandwiched immunocomplex can be formed between the immobilized antibodies on the magnetic beads and the signal antibodies on the gold nanoparticles. The carried DNA initiator strands open the hairpin DNA structures in sequence and propagate a chain reaction of hybridization events between two alternating hairpins to form a nicked double-helix. Numerous ferrocene molecules are formed on the neighboring probe, each of which produces an electrochemical signal within the applied potentials. Under optimal conditions, the immuno-HCR assay presents good electrochemical responses for determination of target IgG at a concentration as low as 0.1 fg mL(-1). Importantly, the methodology can be further extended to the detection of other proteins or biomarkers.
Absorptions by non-phytoplankton particles and phytoplankton, and chromophoric dissolved organic matter (CDOM) were measured at 50 sites in large, shallow, Lake Taihu in winter and summer 2006 to study their seasonal and spatial variations, and their relative contributions to total absorption. The CDOM absorption was significantly higher in winter than in summer, due to degradation and release of fixed carbon in phytoplankton and submerged aquatic vegetation (SAV). The hyperbolic model was used to model the spectral absorption of CDOM, and the mean spectral slope of 6.38 nm -1 was obtained. At most sites, the spectral absorption of non-phytoplankton particles was similar to that of the total particles, demonstrating that the absorption of the total particles is dominated by the absorption of non-phytoplankton particles. In summer, phytoplankton absorption increased markedly, due to frequent algal blooms especially in Meiliang Bay. In winter, the significant increase in non-phytoplankton particle absorption resulted from the increase of inorganic particulate matter caused by sediment resuspension. Strong linear relationships were found between a d (440) and total suspended matter (TSM), organic suspended matter (OSM), and inorganic suspended matter (ISM). Strong linear relationships were also found between a ph (440), a ph (675) and chlorophyll a (Chl-a) concentration. The total relative contributions of non-phytoplankton particles over the range of photosynthetically active radiation (PAR) (400-700 nm) were 48.4 and 79.9% in summer and winter respectively. Non-phytoplankton particle absorption dominated the total absorption, especially in winter, in Lake Taihu, due to frequent sediment resuspension in the large shallow lake as a result of strong windy conditions. The results indicate that strong
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