Fabrication And Application Of Biosensor Chip Based On Chemiluminescence

The microfluidic paper-based devices (μPADs), combining the simplicity of the conventional paper strip tests and the complexity of lab-on-a-chip devices, is a new generation of point-of-care testing and on-site diagnostics. The μPADs, with advantages including low-cost, simple operation, disposable and so on, is well suitable for point-of-care disease diagnostics in developing countries and home health care. Traditional microfluidic devices are made of glass, polymer, silicone, polydimethylsiloxane or plastic, but they are expensive, not disposable and difficult patterning, which limit their extensive application. Paper, which was made of cellulose fibers with high surface area, has the potential to be good alternatives over traditional other materials for the fabrication of the microfluidic devices, because paper is inexpensive, abundant, biodegradable, disposable and easy to modify chemically. The application of the μPADs promotes the rapid development of the microfluidic technology.In this paper, with different fabrication methods, we prepared various of μPADs, based on the advantages of the chemiluminescence analysis including simple equipment, no excitation light source, low background and high sensitivity. In addition, several novel microfluidic paper-based chemiluminescence biosensors were fabricated, and they were applied to the rapid, accurate and specific determination of the target analytes. Finally the novel paper-based chemiluminescence biosensors exhibit acceptable results in real biological samples.The main research in this paper is as follows:1. A novel chemiluminescence system based on M4NRASP and hydrogen peroxide were employed in this work. M4NRASP, a novel rhodanine derivative, was synthetized in our laboratory. The μPADs was prepared by the cutting method, and the urate oxidase was immobilized on the paper surface through simple physical adsorption to fabricate the microfluidic paper-based chemiluminescence enzyme biosensor. It was applied to the rapid, simple, quantitative determination of uric analyte.2. The antibody was immobilized on the paper surface through sodium periodate activating, and the paper zones were prepared by the wax-printing technology. Then the chemiluminescence immunoassay was firstly introduced into the recently proposed μPADs to fabricate the novel paper-based chemiluminescence immunosensors, which has the potential application in point-of-care carcino-embryonic antigen determination. And it was applied in blood samples successfully.3. A novel microfluidic paper-based lab device was fabricated by combining the high sensitivity, high selectivity of the chemiluminescence enzyme-linked immunoassay and the simple operation, low cost of the μPADs. And the chitosan and glutaraldehyde covalent-crosslinking were used to immobilize the antibody. The paper-based device was applied to the simultaneous, rapad determination of multiple tumor markers.4. The capture DNA was immobilized on the μPADs through N,N’-disuccinimidyl carbonate cross-linking to fabricate the novel microfluidic paper-based chemiluminescence DNA biosensor. This biosensor combined the nanoporous gold signal amplification and the chemiluminescence system between carbon dots and potassium permanganate. And it was applied to the rapid, sensitive and trace determination of target DNA.