Study On Mass Spectrometry Rapid Analysis Proteins

Protein is a direct manifestation of life activities,and protein analysis and detection is of great significance to life sciences.The commonly used analytical methods for proteins include enzyme-linked immunosorbent assay,immunoprecipitation,immunoelectrophoresis,and gas chromatography-mass spectrometry.However,these bioassays and chromatographic-mass spectrometry require pretreatment of the sample,which does not allow rapid analysis of the protein.Therefore,it is necessary to establish a real-time direct analysis method.In this paper,paper spray ionization technology and nano-liter electrospray ionization technology are used to establish corresponding methods to analyze and detect proteins.Mainly completed the following contents:(1)Paper spray ionization technology(PSI)is widely used in the analysis of complex samples due to its advantages of simple operation,low consumption and fast analysis speed,it is greatly limited by the hydrogen bond and van der Waals formed between the hydroxyl group on the surface of the paper matrix and the polar functional group in the target compound.In this paper,a new method for preparing hydrophobic paper matrix was developed.The effects of baking temperature and PS microsphere coating amount on the surface properties of impregnated paper were investigated.In this paper,PS impregnated paper as the PSI/MS of matrix,for analysis and detection protein and peptide(e.g.,cytochrome C and lysozyme,myoglobin,angiotensin II and hemoglobin,etc.)in complex matrix(e.g.,PBS,Tris buffer and whole blood samples),compared with filter paper,PS impregnated paper improved the analysis sensitivity of paper spray by 12-1348 times.(2)Metal ions play an important role in living organisms and studying the interaction between metal ions and proteins is an important part of life sciences.In this paper,a method for in situ analysis of metal ions and proteins was established based on Nano ESI technology,and the method was applied to study the interaction between metal ions and proteins.The effects of metal powder type,particle size,metal solution concentration and other factors on the interaction between metal ions and proteins were systematically investigated.The coordination number of the metal ion and the protein is theoretically calculated by the m/z of the characteristic peak of the obtained myoglobin.Finally,the established analytical method is applied to the interaction of other proteins and peptides(such as cytochrome C,lysozyme,angiotensin,etc.)with metal ions,and the coordination number of metal ions and proteins is calculated to be between 1 and 2.The interaction between protein and metal ions was systematically studied,which provided a new idea for protein analysis and detection.