Based On TCM Syndrome Associated With Immune Characteristics Research Of Rare Earth Fluorescent Probe

Objective:According to the urgent needs of precise immunoassay technology in China in recent years,the purpose of this study was to identify the cold coagulation syndrome of acute myocardial infarction（AMI）.A high purity Tb3+hole-like chelating agent satisfying the requirements of homogeneous time resolved fluorescence（HTRF）technology was prepared and successfully labeled troponin antibody.It provides direction and basis for the objective and standardized experimental study of laboratory indicators of TCM syndromes such as acute myocardial infarction cold coagulation heart pulse syndrome.Methods:1.2-benzyloxy lysine-6-tert-butylcarbonylacetic acid E₇was used as the starting material of the reaction,and a polyamino-ligand was synthesized through oxidation,reduction and hydrogenation reactions.All the intermediates were characterized by NMR,IR and MS.2.5-amino-（2-aminoethyl）-H（2,2）-tert-butyl carbamate(E₁₄)was linked to compound B₆（autosynthesis）and modified to synthesize D₂.The fluorescent probe D₃with fluorescence characteristics was synthesized by mixing Tb3+into D₂.D₂and D₃were characterized and confirmed by 1h NMR and IR spectra.The fluorescence spectrum,quantum yield and fluorescence intensity of D₃were studied and analyzed.3.The fluorescent probe D₃was activated by NHS-EDC to label troponin monoclonal antibody K2b5.The activity of labeled antibody and D3 and the performance of fluorescent probe were analyzed.Results:1.Using E₇as raw material,seven intermediates were synthesized.The optimal reaction conditions of E8 and E₉were determined by exploring some reaction conditions.The correctness of the product structure was confirmed by different methods of chromatographic characterization.2.Using E₁₄and B₆as reaction materials,the cave shaped bifunctional chelating agent D₂was synthesized through substitution and structural optimization.The correctness of the product structure was confirmed by spectrometric characterization analysis using different methods.3.The fluorescence probe Tb³⁺lysine-bh（2,2）IMA（D3）was synthesized and the structure of the product was confirmed by spectrogram analysis.By analyzing the quantum yield and fluorescence intensity of D3,it is proved that D3 can be successfully used in HTRF technology.4.By analyzing the activity of labeled antibody and D3 and the performance of fluorescent probe,the results showed that the antibody had good activity before and after labeling,and D3 also maintained good optical properties.It was proved that the self-made fluorescent probe was used to label troponin successfully in this experiment.Conclusion:In this study,the monoclonal antibody complex of troponin labeled with self-made fluorescent probe still maintained immunospecificity and fluorescence performance,providing a technical basis for the study of markers related to myocardial infarction syndrome by HTRF technology.