Synthesis and Characterization of a Novel Europium Chelate: BC-EDTA, and Labeling Nanoparticles for TrFIA

Synthesis and Characterization of a Novel Europium Chelate: BC-EDTA, and Labeling Nanoparticles for TrFIA
　　This dissertation focuses on ultra-high sensitive detection technology. Time-resolved Fluorescence Immunoassay(TRFIA) is such a tool that is applicable to detection of very low concentration of antigen,antibody and pathogenic microorganism.Ball Neodymium Magnets And this dissertation consists of three parts,including a brief review of ultra-high sensitive detection technology,the synthesis and characteration of BC-EDTA;preparation,application of time-resolved fluorescence nanoparticlest and their application in TRFIA.In chapter one,the basic principles and highlights of different kinds of ultra-sensitive detection technology are reviewed.
　　The latest development in TRFIA for protein detection are summarized.And finally the research proposal for this dissertation is presented.In chapter two,we first describe the synthesis of a novel lanthanide chelate:2,9- bis [N,N-bis(carboxymethyl)aminomethyl]-4,7-diphenyl-1,10-phenanthroline(BC-EDTA), which could chelate with Eu3+,and display its fluorescent character.We characterize the chelate by ESI-MS,IR,UV and H1NMR,the results show that the chelate we synthesized is completely correct.In chapter three,we covently lebeled BC-EDTA onto the surface of bare silica nanoparticles,and applied them in TRFIA.The bare silica nanoparticles we synthesized using reverse microemulsion method,and the diameter of particles could be optimized by the ratio of TEOS, http://www.chinamagnets.biz/   H2O and amonia.APTMS was added to introduce amino group onto the surface of bare silica nanoparticles.BC-EDTA can be covently labeled onto the surface of amino-nanoparticles using EDC,NHS in PB(pH=6.8). Finally,amino group was introduced onto the surface of fluorescent nanoparticles, such as antibody,antigen and DNA can be linked on to it.We detected hepatitis B surface antigen using the fluorescenct nanoparticles.The detection limits,calculated with the concentration corresponding to three times of the standard deviations of the background signal,was 65 pg/mL.The upper limit of the calibration curve was ca 200 ng/mL.The coefficient of variation(CV) across the whole range was below 20%.In comparison,under the same conditions,the detection limit of ELISA was 200 pg/mL and the upper limit of the calibration curve was ca 10 ng/mL.