Sodium dodecyl sulfate(SDS) is a powerful solubilizing detergent which is often used during the separation of highly complex protein mixtures by one- or two-dimensional(2D) gel electrophoresis. Matrix-assisted laser d...Sodium dodecyl sulfate(SDS) is a powerful solubilizing detergent which is often used during the separation of highly complex protein mixtures by one- or two-dimensional(2D) gel electrophoresis. Matrix-assisted laser desorption/ionization mass spectrometry(MALDI-MS) is a widely used technique for mass spectrometric analysis of some protein molecules compared to other techniques. But the presence of SDS or some salts usually leads to signal deterioration when using MALDI-MS. A method for using nitrocellulose membrane as the solid-phase carrier combined with n-octyl-β-D-glucopyranoside in the matrix highly enhances the sensitivity of the molecular mass determination of lysozyme. This technique has the advantage that the signal-to-noise of the molecular weight profile is improved compared with the mass spectrum and the profile is relatively easy to interpret.展开更多
A liquid-phase isoelectric focusing electrophoresis system(Rotofor) was used as the prefractionation tool for the sample preparation in the MALDI-MS analysis of a protein mixture. Each fraction collected was then dire...A liquid-phase isoelectric focusing electrophoresis system(Rotofor) was used as the prefractionation tool for the sample preparation in the MALDI-MS analysis of a protein mixture. Each fraction collected was then directly subjected to MALDI-TOF-MS analysis. By this approach, we are able to resolve two types of hemoglobins, A and C, which cannot be successfully separated by means of the traditional SDS-PAGE method.展开更多
A novel sensing chip was designed for MALDI-MS quantitation of acid phosphatase(ACP).The ACP sensing chip was constructed through non-covalent interaction of streptavidin and biotin for the assembly of biotinylated pe...A novel sensing chip was designed for MALDI-MS quantitation of acid phosphatase(ACP).The ACP sensing chip was constructed through non-covalent interaction of streptavidin and biotin for the assembly of biotinylated peptide substrate on biotinylated polyethylene-glycol(PEG)modified indium-tin oxide(ITO)slide.In the presence of ACP,the peptide substrate was dephosphorylated under acidic condition to generate a new mass signal.The quantitative assay of ACP was achieved with the mass signal ratio of product to the sum of product and left peptide substrate.Under optimal detection conditions,the ratio was linearly correlated with the concentration of ACP in the range of 0.05–12 g/L with a detection limit(LOD)of 0.04 g/L.The designed ACP sensing chip has been used to analyze ACP in complex clinical samples,which exhibited high selectivity,good repeatability,and admirably anti-interference ability.This work further demonstrates the concept of MS sensing and the application of MALDI-MS in quantitative analysis,and provides a convenient method for the quantitation of proteases in clinical diagnosis.展开更多
Halogens are elements in soil and water that occur as organic and inorganic species.Studies have shown that inorganic halogens are closely related to animal and plant life.Matrix-assisted laser desorption/ionization m...Halogens are elements in soil and water that occur as organic and inorganic species.Studies have shown that inorganic halogens are closely related to animal and plant life.Matrix-assisted laser desorption/ionization mass spectrometry(MALDI-MS)can achieve high-throughput and rapid analysis,which is also an essential characteristic in some environmental studies.However,there is currently no report on MALDI-MS analysis for water-soluble inorganic halogen compounds.In this study,we aimed to develop a reliable and reproductive method to detect inorganic halogens in soil and water by MALDI-MS.Based on sinapic acid(SA)and silver trifluoromethanesulfonate(SA/AgOTf)as the matrix in negative-ion mode,MALDI-MS showed excellent MS signals related to halogens.Under the optimum conditions,the method achieved satisfactory linearity(R^(2)>0.99)and repeatability(RSD<18%).The LODs for Cl^(−),Br^(−)and I^(−)were 0.76,0.11 and 0.10μg/mL,respectively.The concentration of inorganic halogens from 24 samples was successfully determined by MALDI-MS.In summary,an innovative method was developed for the analysis of inorganic halogens by MALDI-MS using a novel SA/AgOTf matrix,extending the application of MALDI-MS in inorganic compounds.展开更多
提升微生物药敏检测速度是有效救治患者、合理使用抗菌药物、减缓微生物耐药性发展的迫切需要。快速药敏检测的新方法主要是将观察微生物生长的时间点前移,在药敏孵育早期肉眼不可见时,借助不同的设备和技术捕捉微生物数量、形态、代谢...提升微生物药敏检测速度是有效救治患者、合理使用抗菌药物、减缓微生物耐药性发展的迫切需要。快速药敏检测的新方法主要是将观察微生物生长的时间点前移,在药敏孵育早期肉眼不可见时,借助不同的设备和技术捕捉微生物数量、形态、代谢等变化规律,得出微生物与抗菌药物之间的耐受关系。本文从观察微生物的生长现象、荧光标记显示细胞活性、检测细胞代谢状态/产物以及基于基质辅助激光解吸电离飞行时间质谱(matrix⁃assisted laser desorption ionization⁃time of flight mass spectrometry,MALDI⁃TOF MS)的快速药敏4个方面对快速药敏的发展现状进行介绍,以期为临床微生物耐药管理提供参考。展开更多
文摘Sodium dodecyl sulfate(SDS) is a powerful solubilizing detergent which is often used during the separation of highly complex protein mixtures by one- or two-dimensional(2D) gel electrophoresis. Matrix-assisted laser desorption/ionization mass spectrometry(MALDI-MS) is a widely used technique for mass spectrometric analysis of some protein molecules compared to other techniques. But the presence of SDS or some salts usually leads to signal deterioration when using MALDI-MS. A method for using nitrocellulose membrane as the solid-phase carrier combined with n-octyl-β-D-glucopyranoside in the matrix highly enhances the sensitivity of the molecular mass determination of lysozyme. This technique has the advantage that the signal-to-noise of the molecular weight profile is improved compared with the mass spectrum and the profile is relatively easy to interpret.
文摘A liquid-phase isoelectric focusing electrophoresis system(Rotofor) was used as the prefractionation tool for the sample preparation in the MALDI-MS analysis of a protein mixture. Each fraction collected was then directly subjected to MALDI-TOF-MS analysis. By this approach, we are able to resolve two types of hemoglobins, A and C, which cannot be successfully separated by means of the traditional SDS-PAGE method.
基金the National Natural Science Foundation of China(21635005,21827812,21890741,21974063)the National Key Research and Development Program of China(2018YFC1004704)the Fundamental Research Funds for the Central Universities(14380200)。
文摘A novel sensing chip was designed for MALDI-MS quantitation of acid phosphatase(ACP).The ACP sensing chip was constructed through non-covalent interaction of streptavidin and biotin for the assembly of biotinylated peptide substrate on biotinylated polyethylene-glycol(PEG)modified indium-tin oxide(ITO)slide.In the presence of ACP,the peptide substrate was dephosphorylated under acidic condition to generate a new mass signal.The quantitative assay of ACP was achieved with the mass signal ratio of product to the sum of product and left peptide substrate.Under optimal detection conditions,the ratio was linearly correlated with the concentration of ACP in the range of 0.05–12 g/L with a detection limit(LOD)of 0.04 g/L.The designed ACP sensing chip has been used to analyze ACP in complex clinical samples,which exhibited high selectivity,good repeatability,and admirably anti-interference ability.This work further demonstrates the concept of MS sensing and the application of MALDI-MS in quantitative analysis,and provides a convenient method for the quantitation of proteases in clinical diagnosis.
基金This study was jointly supported by the National Natural Science Foundation of China(21906137)the Science and Technology Program of Zhejiang Province(LGC21B050008).
文摘Halogens are elements in soil and water that occur as organic and inorganic species.Studies have shown that inorganic halogens are closely related to animal and plant life.Matrix-assisted laser desorption/ionization mass spectrometry(MALDI-MS)can achieve high-throughput and rapid analysis,which is also an essential characteristic in some environmental studies.However,there is currently no report on MALDI-MS analysis for water-soluble inorganic halogen compounds.In this study,we aimed to develop a reliable and reproductive method to detect inorganic halogens in soil and water by MALDI-MS.Based on sinapic acid(SA)and silver trifluoromethanesulfonate(SA/AgOTf)as the matrix in negative-ion mode,MALDI-MS showed excellent MS signals related to halogens.Under the optimum conditions,the method achieved satisfactory linearity(R^(2)>0.99)and repeatability(RSD<18%).The LODs for Cl^(−),Br^(−)and I^(−)were 0.76,0.11 and 0.10μg/mL,respectively.The concentration of inorganic halogens from 24 samples was successfully determined by MALDI-MS.In summary,an innovative method was developed for the analysis of inorganic halogens by MALDI-MS using a novel SA/AgOTf matrix,extending the application of MALDI-MS in inorganic compounds.
文摘提升微生物药敏检测速度是有效救治患者、合理使用抗菌药物、减缓微生物耐药性发展的迫切需要。快速药敏检测的新方法主要是将观察微生物生长的时间点前移,在药敏孵育早期肉眼不可见时,借助不同的设备和技术捕捉微生物数量、形态、代谢等变化规律,得出微生物与抗菌药物之间的耐受关系。本文从观察微生物的生长现象、荧光标记显示细胞活性、检测细胞代谢状态/产物以及基于基质辅助激光解吸电离飞行时间质谱(matrix⁃assisted laser desorption ionization⁃time of flight mass spectrometry,MALDI⁃TOF MS)的快速药敏4个方面对快速药敏的发展现状进行介绍,以期为临床微生物耐药管理提供参考。
