| Based on new type of magnetic supporting material ?MWCNTs@Fe3O4@SiO2, magnetic SiO2/MWCNTs,?-cyclodextrin/chitosan-magnetic graphene oxide and ionic liquid modified magnetic graphene oxide//?-cyclodextrin?, surface molecular imprinting polymer ?SMIP? with specially recognition properties was obtained. Combined with Chemiluminescence ?CL? analysis techniques, SMIP-CL biosensor was achieved. Simultaneously, different simple easy combination way was also designed. By avoiding the traditional complex sample pretreatment process, the proposed biosensor could do the control analysis in real-time with high sensitivity and selectivity, and the results was satisfactory.In the first part, an ultrasensitive CL biosensor with high selectivity based on bioreceptor SMIP which using core-shell Fe3O4@SiO2-multi-walled carbon nanotubes nanostructures ?Fe3O4@SiO2@MWCNTs? as backbone material for bovine hemoglobin ?BHb? was proposed. The Fe3O4@SiO2@MWCNTs was synthesized with a new method, and then was characterized by SEM, FTIR and XRD techniques. Adsorption ability of the Fe3O4@SiO2@MWCNTs-SMIP was evaluated to be 91 mg/g and Fe3O4@SiO2@MWCNTs-SMIP followed Langmuir isotherm equation and it demonstrated the excellent recognition and adsorption ability for the imprinted cites which located on the surface or near the surface of the Fe3O4@SiO2@MWCNTs. Under the optimum conditions of CL, the detection range of BHb was from 5.0 × 10-10 to 7.0 × 10-7 mg/mL with the detection limit of 1.5 × 10-1 mg/mL ?3d?. The proposed biosensor was successfully applied in determination of BHb in real samples with high selectivity and sensitivity, and the recoveries were excellent and varied from 92% to 106%. Finally, the possible CL mechanism of the BHb amplifying the CL signal of luminol-NaOH-H2O2 system was discussed.In the second part, a CL sensor with high sensitivity and selectivity has been developed for the determination of Ribonuclease A ?RNase A?. A new material FeO4/multi-walled carbon nanotubes/SiO2 ?Fe3O4/MWCNTs/SiO2? was introduced into this sensor as supporting material to prepare SMIP. In this work, Fe3O4 could not only serve as backbone material in the preparation of RNase A SMIP, but also as separation reagent to make the collection of SMIP complex easily, carbon nanotube and SiO2 was used as supporting material to bear SMIP for their large specific surface area. Then, the nanocomposite of Fe3O/MWCNTs/SiO2 was characterized by scanning electron microscopy ?SEM?, X-ray diffraction ?XRD? and Fourier transform infrared spectroscopy ?FT-IR? techniques. The adsorption ability of Fe3O4/MWCNTs/SiO2-SMIP was calculated to be 102 mg/g and it demonstrated the excellent recognition and adsorption ability of the imprinting cavities situated at or in the proximity of the surface of the Fe3O4/MWCNTs/SiO2. Under the optimal conditions, the linear range extended from 1.0 x 10-9 mg/mL to 1.0 x 10-7 mg/mL for RNase A and the detection limit was 3.2×10-10 mg/mL ?3S?. The proposed sensor was successfully applied in determination of RNase A in biological samples with recoveries from 93% to 105%.In the third part, a sensitive and selective CL biosensor for bovine serum albumin ?BSA? coupled with surface molecularly imprinted polymer nanocomplex using ^-cyclodextrin/chitosan-magnetic graphene oxide as backbone material ??-CD/Cs-MGO-SMIP? was promoted. Then, the material ?-CD/Cs-MGO in which ?-cyclodextrin, chitosan and graphene oxide was used to provide multi-imprinting sites and large surface area was characterized by SEM, XRD and FTIR, and then it was found that ?-CD/Cs-MGO-SMIP followed Langmuir isotherm equation and pseudo-second order sorption kinetics when binding the template. It demonstrated fast mass transfer, promoted rate of removal of the biomacromolecule and excellent recognition and adsorption ability for the imprinting cavities situated at the surface of th ?-CD/Cs-MGO, which enabled easy access to BSA. Subsequently, a high sensitive CL biosensor to BSA based on the strong recognition effect between ?-CD/Cs-MGO-SMIP and BSA which decided the high selectivity has been proposed and the proposed biosensor could be assay in the range of 5.0 x 10-7-1.0×10-4 mg/mL with a detection limit of 1.1×10-7 mg/mL. The obtained recoveries were between 94% and 106% when determining samples.In the fourth part, ionic liquid modified Fe3O4@dopamine/graphene oxide/ ?-cyclodextrin ?ILs-Fe3O4@DA/GO/?-CD? was used as supporting material to synthesize SMIP which then was introduced into CL to achieve an ultrasensitive and selective biosensor for determination of lysozyme ?Lys?. ILs and ?-CD was applied to provide multiple binding sites to prepare Lys SMIP and Fe3O4@DA was designed to make the product separate easily and prevent the aggregation of GO which could improve absorption capacity for its large specific surface area. The ILs-Fe3O4@DA/GO/?-CD-SMIP showed high adsorption capacity ?Q=101 mg/g? to Lys in the adsorption isotherm assays. The adsorption equilibrium was reached within 10 min for all the concentrations, attributing to the binding sites situated exclusively at the surface, and the adsorption model followed Langmuir isotherm. Under the suitable CL conditions, the proposed biosensor could response Lys linearly in the range of 1.0 × 10-9-8.0 × 10-8 mg/mL with a detection limit of 3.0× 10-10 mg/mL. When used in practical samples in determination of Lys, the efficient biosensor exhibited excellent result with the recoveries ranging from 94% to112%. |
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