| The research work of this thesis is divided into two parts: First of all,in view of the existing adenosine detection methods,which is constructed with complex pretreatment,time-consuming operation,expensive equipment and difficult to popularize.We invented a new adenosine detection system aimed at achieving those purposes that low cost,simple operation,rapid detection,easy access to the instrument,and can be used to achieve specific and accurate quantitative analysis of adenosine in biological samples.And intend to combine with the second part of the study for achieving the detection of adenosine quantitative which is enrichment by adenosine molecularly imprinted polymer microspheres;Secondly,a simple and effective method for the preparation of core-loaded adenosine molecularly imprinted polymer microspheres(MIP)was proposed,and the MIP prepared is not only specifically recognized for adenosine,but also for ADP-ribosylated protein can exhibit good selectivity.The research work of this thesis mainly includes the following contents:(1)A new method for the detection of adenosine based on TPE and ABA was constructed by combining the aggregation-induced luminescence properties possessed by TPE with the specific recognition properties of the target substance by the nucleic acid aptamer(ABA).Then the optimal concentration of TPE/ABA was detected as 10:0.1,and the lowest detection concentration of adenosine in the detection system was 10 pM,through the fluorescence detection of adenosine and its three analogues(cytidine,uracil,guanine),it was found that this detection method had a good selectivity for adenosine,at the same time,the recovery rate was 86.8% ~ 90.0%,which proved that the method has good precision and accurate and reliable in urine samples.(2)Polystyrene microspheres(CP)with RAFT chain transfer groups were prepared by the combination of traditional free radical precipitation polymerization and reversible addition-fracture chain transfer polymerization(RAFT).Three groups of parallel samples were prepared under certain reaction conditions(reaction temperature: 70°C,polymerization time: 3.5 h),and then the yield and appearance of the three groups were compared.The experimental results show that the yield,particle size and appearance of CP microspheres which is synthesized by RAFTPP under the same conditions appearance are more stable.(3)Utilizing the RAFT chain transfer group on the surface of the CP microspheres,the second part of the polymerization reaction can be carried out again with adenosine as the template molecule,so that MIP microspheres with hydrophilic properties on the surface of the microspheres were prepared by forming a layer of molecular imprinted shells on the outer surface of the microspheres.And the yield,particle size and dispersibility of MIP microspheres prepared at different temperature and time were compared and analyzed to determine the optimum reaction temperature was 70℃ and reaction time 60 min.(4)The CP,MIP and NIP were characterized by scanning electron microscopy,water suspension test,adenosine reabsorption test and protein adsorption experiment.The results show that the MIP prepared by RAFTPP method has better dispersion stability in water,and the molecular recognition performance of MIP in biological solution can be improved by the hydrophilicity of MIP molecularly imprinted shells.There is no obvious distinguish of the reabsorption of adenosine between the three microspheres,what’s more,both MIP and NIP all had a lot of adsorption on the gapA protein but the adsorption effect was not significantly different.The main reason for this phenomenon may be that the adenosine imprinting was unsuccessful in the preparation of MIP,the number of adenosine footprints on its surface is too small or the cavities are problematic so that adenosine molecules can’t be recognized. |
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