Studies On The Chemiluminescence Immunolabelling Technique Based On Magnetic Beads And Its Applications

This paper discussed the novel and sensitive chemiluminescence immunoassays integrating with the magnetic beads technique, labelling technique and nanotechnique. The CL reaction conditions and the performance of the biosensors were studied by using chemiluminescence assay and spectral analysis. In summary, the high sensitivity, ease of fabrication and a low cost make methods in this paper may provide an interesting alternative tool for detection protein in clinical laboratory. And it has a good developmental prospect.The main jobs of this paper can be concluded as follows:1. The study on the CL magnetic enzyme-linked immunoassay and its applicationA CL magnetic enzyme-linked immunoassay was developed integrating with the magnetic beads and sandwich immunocomplexes. The concentration ofα-fetoprotein (AFP) was monitored based on the HRP label activity toward the oxidation of luminol, which was quantified by CL method. The linear range for human AFP was 0.8 ng/mL -8.0 ng/mL. The CL magnetic enzyme-linked immunoassay for clinical applications was investigated by analyzing several real samples, in comparison with the ELISA method. The calibration curve indicated that there was no significant difference between the results given by two methods.2. The study on the CL magnetic enzyme-linked immunoassay based on luminol/antibody labeled gold nanoparticles and its applicationA facile magnetic enzyme-linked immunoassay by loading luminol and secondary antibody on gold nanoparticles (Au NPs) was described in this work. The label method was further evaluated via a sandwich-type CL immunoassay of CEA in serum. In this protocol, the CEA analyte was captured by the primary antibody immobilized on the surface of magnetic beads, and then was sandwiched by the secondary antibody loaded on luminol-labeled Au NPs. The linear range for CEA was 5.0×10-9-2.0×10-8 g/mL by using Co2+ as catalysts. The CL magnetic enzyme-linked immunoassay based on luminol/antibody labeled gold nanoparticles for clinical applications was investigated by analyzing several real samples, in comparison with the ELISA method. The calibration curve indicated that there was no significant difference between the results given by two methods.3. The study on the CL detection of GSH using Isoluminol labeled on polystyrene microbeadsA new strategy was designed for detecting GSH and free thiols in K562 cells by chemiluminescent system of Isoluminol-H2O2-HRP through GSH cleaving disulfide. Chemiluminescence detection is of high sensitivity, rapidity and simplicity and the linear range of GSH was 5×10-10 to 8×10-8 M. This strategy presents a new method to determine GSH and thiols with high veracity and feasibility.