Development Of An Electrochemical Multiresidue Immunosensor For Advanced Glycation End Products

Advanced glycation end products(AGEs)are irreversible polymers that they are products of a series of complex non-enzymatic glycosylation reactions in biological macromolecules such as nucleic acids and proteins,and they are closely related to the occurrence and development of various diseases in the human body,it has practical effects on the prevention and treatment of related diseases and the monitoring of the degree of disease to realize the routine clinical detection of AGEs.However,because of different species,complex structures and undefined structures of the AGEs,the mothods used todayin medical and academic research communities fail to meet the requirements actual clinical detection of AGEs.Therefore,designing and developing an efficient,low-cost,clinically-applicable,innovative technology for multi-residue detection of AGEs is an urgent problem to be solved.In this study,three different kinds of novel AGEs electrochemical multi-residue immunosensors were constructed in combination with anti-pentosidine monoclonal antibodies.The highly sensitive and multi-residue detection of AGEs was achieved,these immunosensors are turned out very well when applied to detection AGEs in real serum samples.Chapter 1:The production mechanism,structure,pathogenic mechanism,and biochemical properties of AGEs are introduced.We make the summary of current status of AGEs research,common detection methods and their advantages and disadvantages in this chapter.Last the thesis selection basis and innovation are introduced.Chapter 2:The graphene-chitosan composite was successfully prepared and characterized by ultraviolet-visible absorption spectroscopy,laser Raman spectroscopy and scanning electron microscopy,anti-pentosidine monoclonal antibody was prepared,Ns-(carboxymethyl)-lysine,Nε-(carboxyethyl)-lysine,Argpyrimidine,MG-Imidazolones four AGEs molecules were prepared by organic synthesis.To construct a BSA/pentosidine/graphene-chitosan electrochemical biosensor for the detection of pentosidine and four synthetic AGEs.The relationship curve,linear range,and minimum detection limit of the five kinds of AGEs were determined,also the selectivity,stability,and reproducibility were studied.Then the actual serum samples were tested for spike recovery,and the average recoveries were in the range of 84.45-107.4%.Chapter 3:The Nε-(carboxymethyl)-lysine/thioglycolic acid/nanogolds/GCE electrochemical biosensor was constructed by chemical reaction,and infrared spectroscopy was used to characterize the construction process.Then,Nε-(carboxymethyl)-lysine,Nε-(carboxyethyl)-lysine,pentosidine,Argpyrimidine,and MG-Imidazolones were detected using the built-in sensors.The relationship curve,linear range,and minimum detection limit of the five kinds of AGEs were determined,and the selectivity,stability,and reproducibility were studied.Then the actual serum samples were tested for spike recovery,and the average recoveries were in the range of 96.90-107.03%.Chapter 4:Gold nanowire electrodes were successfully prepared and characterized by transmission electron microscopy.The Nε-(carboxymethyl)-lysine/thioglycolic acid/gold nanowire electrochemical biosensor was constructed by chemical bond linkage to detect five ultra-low concentrations of AGEs.The relationship curve,linear range,and minimum detection limit of the five kinds of AGEs were determined,finally the selectivity,stability,and reproducibility were studied.