Application Of Novel Nanozymes In Biological Analysis

Nanozyme is a kind of nano-scale material that has intrinsic enzymatic activity and it can be used as a substitute for natural enzymes with the advantages of stability,low cost and recyclability.Among them,nanozymes with peroxidase-like and oxidase-like properties have attracted wide attention.Because,in the presence or absence of hydrogen peroxide,it can catalyze the color change of the enzyme substrate.Through this color reaction,people can adopt a simple,fast and practical colorimetric method to achieve the sensitive detection of biomolecules.In this paper,a variety of nanomaterials with different morphology were designed and synthesized.Their peroxidase or oxidase-like properties were analyzed in detail.We use these properties to achieve the detection of glucose molecules and uric acid molecules.Nanozymes have great application potential in biological analysis through our research work.?1?In this research work,we used sulfur-containing organic matter glutathione?GSH?to provide sulfur,combing with the reducing properties of GSH,and simply and quickly synthesized inorganic FeS nanosheets.The raw materials used in FeS nanosheets are simple and the synthesis steps are easy to operate,which can greatly reduce production cost.It has the inherited peroxidase activity and can catalyze the peroxidase substrates 3,3',5,5'-tetramethylbenzidine?TMB?and hydrogen peroxide?H₂O₂?.A steady-state kinetics experimental study was conducted and we found that its affinity for substrates is similar to Horse Radish Peroxidase?HRP?.it can replace HRP for molecular detection and reduce the preparation cost.Combining FeS nanozyme with colorimetric method,we successfully detect the glucose.The linear range of this method is 1-150?M and the limit of detection is as low as0.3?M.The method has strong anti-interference ability and is successfully applied to the detection of human blood glucose.?2?In this study,we successfully prepared PtBi nanosheets with a triangular structure,which is a new type of nanozyme.Compared with the single metal Pt,it not only has the peroxidase-like activity of Pt,but also enhanced the peroxidase activity of Pt.Its activity can be adjusted by adjusting the added amount of Bi.Compared with HPR,it has better temperature tolerance and better affinity for TMB.We use the excellent properties of PtBi nanozyme connecting with the colorimetric method and apply it in the detection of glucose with good detecting results.The linear range of glucose detection is 0.1-300?M,the limit of detection is as low as 0.05?M.This method has strong selectivity for glucose and realizes the detection of human blood glucose.?3?In this research work,we used a simple and rapid synthesis method to prepare CoOOH nanosheets and reported the material owned good oxidase-like activity for the first time.It can directly oxidize colorless TMB to its corresponding blue color oxidation state?ox TMB?.Based on CoOOH nanozyme,the substrate of TMB was firstly oxidized into ox TMB,and then added uric acid with reducing properties,the color became lighter,and at the same time the absorbance was also changed,so we successfully achieved the detection of uric acid.The linear range is 5-150?M,and the limit of detection is 1.6?M,which is to meet the needs of clinical testing.