An ultra-facile and label-free immunoassay strategy for detection of copper (II) utilizing chemiluminescence self-enhancement of Cu (II)-ethylenediaminetetraacetate chelate.

The establishment of facile, rapid, sensitive and cost-effective protocols for the detection of heavy metals is of great significance for human health and environmental monitoring. Hereby, an ultra-facile and label-free immunoassay strategy was designed for detecting heavy metal ion by using Cu (II) as the model analyte. Cu (II) reacted previously with ethylenediaminetetraacetate (EDTA) was captured by immobilized monoclonal antibody for Cu (II)-EDTA chelate. Then Cu (II) was detected based on the self-enhancing effect of Cu (II)-EDTA chelate to luminol-H2O2 chemiluminescence reaction. The CL intensity is linear relative with Cu (II) concentration in a very wide range of 1.0-1000ng/mL, with a detection limit of 0.33ng/mL (S/N=3). Since the specificity of this proposed strategy relied on both the specificity of monoclonal antibody and the specificity of luminol-H2O2 system, it could avoid interference from most common ions. The proposed method was used successfully to detect Cu (II) in traditional Chinese medicine and environmental water samples with acceptable recovery values of 82-113%. This proof-of-principle work demonstrated the feasibility of the label-free immunoassay for heavy metal ions, and opened a new avenue for rapid screening and field assay for drug safety, environmental monitoring and clinical diagnosis.

Chemiluminescence reaction kinetics-resolved multianalyte immunoassay strategy using a bispecific monoclonal antibody as the unique recognition reagent.

The multianalyte immunoassay (MIA) has attracted increasing attention due to its high sample throughput, short assay time, low sample consumption, and reduced overall cost. However, up to now, the reported MIA methods commonly require multiple antibodies since each antibody can recognize only one antigen. Herein, a novel bispecific monoclonal antibody (BsMcAb) that could bind methyl parathion and imidacloprid simultaneously was produced by a hybrid hybridomas strategy. A chemiluminescence (CL) reaction kinetics-resolved strategy was designed for MIA of methyl parathion and imidacloprid using the BsMcAb as the unique recognition reagent. Horseradish peroxidase (HRP) and alkaline phosphatase (ALP) were adopted as the signal probes to tag the haptens of the two pesticides due to their very different CL kinetic characteristics. After competitive immunoreactions, the HRP-tagged methyl parathion hapten and the ALP-tagged imidacloprid hapten were simultaneously bound to the BsMcAb since there were two different antigen-binding sites in it. Then, two CL reactions were simultaneously triggered by adding the CL coreactants, and the signals for methyl parathion and imidacloprid detections were collected at 0.6 and 1000 s, respectively. The linear ranges for methyl parathion and imidacloprid were both 1.0-500 ng/mL, with detection limits of 0.33 ng/mL (S/N = 3). The proposed method was successfully used to detect pesticides spiked in ginseng and American ginseng with acceptable recoveries of 80-118%. This proof-of-principle work demonstrated the feasibility of MIA using only one antibody.

[Effects of nighttime warming on winter wheat root growth and soil nutrient availability].

Climate warming has an obvious asymmetry between day and night, with a greater increment of air temperature at nighttime than at daytime. By adopting passive nighttime warming (PNW) system, a two-year field experiment of nighttime warming was conducted in the main production areas of winter wheat in China (Shijiazhuang of Hebei Province, Xuzhou of Jiangsu Province, Xuchang of Henan Province, and Zhenjiang of Jiangsu Province) in 2009 and 2010, with the responses of soil pH and available nutrient contents during the whole growth periods and of wheat root characteristics at heading stage determined. As compared with the control (no nighttime warming), nighttime warming decreased the soil pH and available nutrient contents significantly, and increased the root dry mass and root/shoot ratio to a certain extent. During the whole growth period of winter wheat, nighttime warming decreased the soil pH in Shijiazhuang, Xuzhou, Xuchang, and Zhenjiang averagely by 0.4%, 0.4%, 0.7%, and 0.9%, the soil alkaline nitrogen content averagely by 8.1%, 8.1%, 7.1%, and 6.0%, the soil available phosphorus content averagely by 15.7%, 12.1%, 19.6%, and 25.8%, and the soil available potassium content averagely by 11.5%, 7.6%, 7.6% , and 10.1%, respectively. However, nighttime warming increased the wheat root dry mass at heading stage in Shijiazhuang, Xuzhou, and Zhenjiang averagely by 31. 5% , 27.0%, and 14.5%, and the root/shoot ratio at heading stage in Shijiazhuang, Xuchang, and Zhenjiang averagely by 23.8%, 13.7% and 9.7%, respectively. Our results indicated that nighttime warming could affect the soil nutrient supply and winter wheat growth via affecting the soil chemical properties.