Research On New Method Of Design And Making Of Immune Paper Chip

Paper-based microfluidic devices have recently gained significant interest because of their advantages of rapid, low-cost, miniaturization portable and easy to operate. The main purpose of this thesis was to combine the paper-based chip with immunoassay, seeking a novel washing strategy, finding a new method to fabricate the paper-based chip. The thesis is divided into two parts, the first part is the review, which summarized the property, fabrication method and application of the paper-based chip. The second part is as follows:1. A new fabrication method of paper-based analytical device integrated with multiplexed chemiluminescence immunodeviceA multiplexed chemiluminescence immunodevice was developed for fast and sensitive determination of carcinoembryonic antigen (CEA), a-fetoprotein (AFP), prostate-specific antigen (PSA) simultaneously, incorporated into a low-cost microfluidic paper-based analytical device (μPAD). The μPAD was fabricated by hydrophobic reagent dipping method, which was comprised of three detection zones, one reagent inject zone and three flow channels. The hydrophobic reagent was formed by dissolving the recycling foamed plastic in chloroform. Chitosan coating and glutaraldehyde cross-linking method was used to modify μPAD to covalently immobilize antibodies on μPAD. After sandwich-type immunoreactions, the HRP labeled antibody was captured on the surface of μPAD to catalyze the luminol-H2O2 CL system upon addition of p-iodophenol,which produced an enhanced CL emission. Using CEA, AFP, PSA as model analytes, this approach provided good linear range from 0.1 to 80 ng/mL,5 to 80 ng/mL,1 to 50 ng/mL respectively with low detection limits of 0.05 ng/mL,1.5 ng/mL,0.3 ng/mL respectively under optimal conditions. Accurate determinations of these proteins in human plasma samples were demonstrated by comparison to the standard ELISA method. This paper-based microfluidic CL detection system provides a new strategy for a low-cost, sensitive, simultaneous multiplex immunoassay.2. A novel washing technique integrated paper-based microfluidic immunodeviceA novel washing strategy with a ring-oven integrated on the paper-based microfluidic immunodevice for the detection of carcinoembryonic antigen (CEA) is presented in this work. The washing procedure of the immunoassay can be achieved well with the wasting area heated on the paper device by putting the ring-oven under the paper. The washing solution diffused to the washing area freely by the capillary force and then dried by the ring-oven which can precisely control the temperature of the different part on the paper-based chip. The paper device which was composed of eight microfluidic channels was fabricated by a low-cost, simple, and rapid paper cutting method. Chitosan coating and glutaraldehyde cross-linking were used to modify μPADs to covalently immobilize antibodies on μPADs. Lower detection limit of 0.03 ng/mL CEA can be obtained by enzyme-linked immunosorbent assay (ELISA). The washing efficiency was increased greatly comparing to the traditional washing methods and the established paper-based device can be used in the determination of CEA in human serum with higher sensitivity. The paper-based device provides a new washing strategy for sensitive immunoassay and point-of-care diagnostics.3. A paper-based chemiluminescence device for the determination of ofloxacinIn this work, a wax-printed paper-based analytical device combined with silver nanoparticles (AgNPs) catalyzed luminol chemiluminescence (CL) system for the determination of ofloxacin (OFLX) was presented. It was based on the enhancement of CL intensity of luminol-H2O2 system by AgNPs. Under the selected experimental conditions, a linear relationship was obtained between the CL intensity and the concentration of OFLX in the range from 1.0×10-9 g/mL to 1.0×10-6 g/mL with a detection limit of 3.0×10-10 g/mL. This study shows the successful integration of the paper-based chip with the CL method. It will be of interest for use in areas such as disease diagnosis in the future.