Goldmag Nanocomposites And Fluorescent Nanoparticles Based Sandwich Assay For Myoglobin Detection

Immunoassay technique, which is based on antigen-antibody interaction, has been applied for a broad range of applications, such as clinical diagnosis, food safety, and environmental analysis. This research is to introduce GoldMag nanocomposites and fluorescent nanoparticles into immunoassay and to make a exploration of nanoparticles applications for myoglobin detection. So three part of work have been done in this paper, as follow:1. Fe3O4 magnetic nanoparticles with diameter around of 150 nm were prepared by solvothermal method, then MNPs were coated by silica and carboxyl modified by APTES-COOH. Follow by BSA decoration and gold nanoparticle loading, finally, the MNP@SiO2@BSA@Au nanocomposites were prepared. The TEM results showed gold nanoparticle with diameter of around 15 nm were loaded on the MNP@SiO2 surface and the particle dispersity of MNP@SiO2@BSA@Au was good.2.Three kinds of fluorescent nanoparticles were prepared, SNP@Rh X, PAAAM@Rh X and SNP@PAAAM@Rh X nanoparticles. SNPs were synthesized by St?ber method, then incorporated with Rh X and carboxyl modified by APTES-COOH, finally SNP@Rh X nanoparticles were prepared. PAAAM nanoparticles were prepared by reversed-phase microemulsion method and then Rh X was coupled to PAAAM nanoparticle s(PAAAM@Rh). And for SNP@ PAAAM@Rh X nanoparticles preparation, SNPs were firstly modified by(3-Mercaptopropyl)trimethoxysilane( MPS), then a layer of PAAAM was polymerized on the surface of SNPs, follow by Rh X conjugation. The TEM results indicated SNP@Rh X and SNP@PAAAM@Rh X nanoparticles had a good particle dispersity.3.This research established a new detection method for myoglobin based on sandwich assay. In the present of myoglobin, magnetic nanoparticles bearing 4E2 antibody and fluorescent nanoparticles bearing 7C3 antibody can form a sandwich structure,which can be separated from blood under magnetic field. The concentration of myoglobin based on fluorescence intensity of SNP@Rh X nanoparticles in the sandwich structure. The calibration curve of the myoglobin showed the linear range between 0~250 ng/mL(R2 = 0.99). The minimum detectable concentration was 0.28 ng/mL. The fluorescence method offered a simple and quick way to determine myoglobin with a total analysis time of less than 30 min in human blood.