Study On C - Reactive Protein Immunoassay Based On Metal - Organic Skeleton Materials And

Metal-organic framework materials(MOFs), with the advantages of porous structure, large specific surface area and easy modification, have attracted many researchers. SNHKUST-1, a kind of Cu-MOF, has the characteristics of nanomimices. In this thesis, a series of nanomimices with good biocompatibility and strong catalytic activity, such as hollow silver platinum nanoparticles(Ag/Pt), Au / SNHKUST-1 Pt / SNHKUST-1, Pt / MnO2 nanocomposite, was synthesized and used to construct immunosensors for the determination of C- reactive protein(CRP). Details are as follows:(1) Hollow silver platinum nano-materials was synthesized and characterized using displacement reaction hydrothermal synthesis method. A sensitive sandwich C- reactive protein immunosensor was prepared by means of the catalytic activity of Ag/Pt to the reduction of H2O2. Firstly, antibody of C- reactive protein was immobilized on the electrode through hollow graphene material. Secondary antibody of C- reactive protein was labeled with hollow Ag/Pt nanoparticles and C- reactive protein was detected by a sandwich method. Under the optimal experimental conditions, the linear range of C- reactive protein is 0.1 ~ 150 ng / mL with linear correlation coefficient of 0.9643 and the detection limit of 0.02 ng / mL.(2) Au nanoparticles-loaded metal organic framework compounds with ultranano-structured(Au/SNHKUST-1) were synthesized at room temperature and used as a tag to label anti C-creative protein. Hollow graphene nanomaterials doped with conductive material Au nanoparticles were also synthesized and utilized as a immobilized matrix through π-π interact absorption between hollow graphene and glassy carbon electrode to enhance the amount of the immobilized antibody. The quantitative determination of C- reactive protein can be achieved by detecting the catalytic current of label to the reduction of H2O2. The response current has a good linear relationship with C- reactive protein concentration within 0.2 ng / mL-200 ng / mL with linear correlation coefficient of 0.9947 and the detection limit of 0.05 ng / mL. This immunosensor of C- reactive protein provides a new simple and rapid detection method for CRP.(3) A kind of Cu-MOF nanoparticles(SNHKUST-1) were synthesized and loaded with Pt nanoparticles, which was used as a label to construct sandwich immunosensor of C- reactive protein using synthesized graphene oxide as a immobilized matrix.The quantitative determination of C- reactive protein can be achieved by detecting the catalytic current to the reduction of H2O2 of this label. The linear range of this sensor was 0.1ng / mL-100 ng / mL. The linear equation was I = 0.72894 C + 1.43293 with the detection limit of 0.033 ng / mL and the linear correlation coefficient R2 was 0.9915. This immunosensor can be used to detect CRP in serum sample.(4) Enzyme linked immunosorbent assay(ELISA) is a method of combining antibody with enzyme complex, and then to detect antigen by color. In this chapter,Pt / MnO2 nanocomposite were synthesized and bound with glucose oxidase at room temperature, which was used to label CRP antibody. Glucose oxidase(GOx) has the specific catalytic activity to β-D(+) glucose and produce an intermediate product H2O2, which can have chromogenic reaction with TMB(3,3’,5,5’-Tetramethylbenzidine). In this study, the concentration of CRP was determined by sandwich ELISA using UV-visible spectrophotometer to detect absorbance which is proportional to the concentration of CRP within 0.05ng/mL-2000.00 ng/mL under optimal conditions. This method can be used for the quantitative determination of CRP in real sample.