• 제목/요약/키워드: Matrix-assisted laser desorption mass spectrometry

검색결과 202건 처리시간 0.023초

Fast visible dye staining of proteins in one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gels compatible with matrix-assisted laser desorption/ionization-mass spectrometry

  • Jin, Li-Tai;Hwang, Sun-Young;Yoo, Gyurng-Soo;Choi, Jung-Kap
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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    • pp.147.2-147.2
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    • 2003
  • A fast and matrix-assisted laser desorption/ionization-mass spectrometry compatible protein staining method in one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis is described. It is based on the counterion dye staining method that employs oppositely charged two dyes, Zincon and Ethyl Violet to form an ion-pair complex. It is safe to use since the methanol used previously in staining solution was replaced with ethanol, which is not toxic. The protocol including fixing, staining and quick washing steps can be completed in 1 to 1.5 h depending upon gel thickness. (omitted)

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Novel analysis procedure for red ginseng polysaccharides by matrix-assisted laser desorption/ionization time-of-flight/time-offlight mass spectrometry

  • Jin, Ye Rin;Oh, Myung Jin;Yuk, Heung Joo;An, Hyun Joo;Kim, Dong Seon
    • Journal of Ginseng Research
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    • 제45권5호
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    • pp.539-545
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    • 2021
  • Background: Red ginseng polysaccharides (RGPs) have been acknowledged for their outstanding immunomodulation and anti-tumor activities. However, their studies are still limited by the complexity of their structural features, the absence of purification and enrichment methods, and the rarity of the analytical instruments that apply to the analysis of such macromolecules. Thus, this study is an attempt to establish a new mass spectrometry (MS)-based analysis procedure for RGPs. Methods: Saponin pre-excluded powder of RG (RG-SPEP, 10 mg) was treated with 200 µL of distilled water and centrifuged for 5 h at 1000 rpm and 85 ℃. Ethanol-based precipitation and centrifugation were applied to obtain RGPs from the heated extracts. Further, endo-carbohydrase treatments were performed to produce specific saccharide fragments. Solid-phase extraction (SPE) processes were implemented to purify and enrich the enzyme-treated RGPs, while matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) MS was employed for the partial structural analysis of the obtained RGPs. Results: Utilizing cellulase, porous graphitized carbon (PGC), hydrophilic interaction chromatography (HILIC), and MALDI-TOF/TOF MS, the neutral and acidic RGPs were qualitatively analyzed. Hexn and Hexn-18 (cellulose analogs) were determined to be novel neutral RGPs. Additionally, the [Unknown + Hexn] species were also determined as new acidic RGPs. Furthermore, HexAn (H) was determined as another form of the acidic RGPs. Conclusion: Compared to the previous methods of analysis, these unprecedented applications of HILIC-SPE and MALDI-TOF/TOF MS to analyze RGPs proved to be fairly effective for fractionating and detecting neutral and acidic components. This new procedure exhibits great potential as a specific tool for searching and determining various polysaccharides in many herbal medicines.

Construction of an Improved Tandem Time-of-flight Mass Spectrometer for Photodissociation of Ions Generated by Matrix-assisted Laser Desorption Ionization (MALDI)

  • Moon, Jeong-Hee;Yoon, So-Hee;Kim, Myung-Soo
    • Bulletin of the Korean Chemical Society
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    • 제26권5호
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    • pp.763-768
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    • 2005
  • An improved tandem time-of-flight (TOF) mass spectrometer for the photodissociation (PD) study of ions generated by matrix-assisted laser desorption ionization (MALDI), MALDI-TOF-PD-TOF, has been designed and constructed. Recording a full spectrum with better than unit mass resolution even in low mass range has been achieved without reflectron voltage stepping which was needed in the previous version. Other aspects of the improvement, such as those in the data system which now allow 10-100 times faster spectral acquisition than with the previous instrument, are described. Rationale for the ideas which have led to the improvements is presented also.

Enrichment of Peptides using Novel C8-functionalized Magnetic Nanoparticles for Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometric Analysis

  • Song, Sun-Mi;Yang, Hyo-Jik;Kim, Jin-Hee;Shin, Seong-Jae;Park, Eun-Hye;Kim, Jeong-Kwon
    • Mass Spectrometry Letters
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    • 제2권2호
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    • pp.53-56
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    • 2011
  • [ $C_8$ ]functionalized magnetic nanoparticles were synthesized by coating magnetic $Fe_3O_4$ nanoparticles with silicaamine groups using 3-aminopropyltriethoxysilane and by subsequently modifying the amine groups with chloro(dimethyl)octylsilane to produce octyl groups on the surface of the MNPs. The $C_8$-functionalized MNPs were used to enrich peptides from tryptic protein digests of myoglobin and ${\alpha}$-casein. The enriched peptides were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). MALDI-MS was also used to investigate desalting of the $C_8$-functionalized MNPs. Sample solutions were prepared in 1.0 M NaCl, and the successful removal of salt was observed. Enrichment with $C_8$-functionalized MNPs was very effective for separating and concentrating tryptic peptides.

Visible Surface-Assisted Laser Desorption/Ionization Mass Spectrometry of Small Macromolecules Deposited on the Graphite Plate

  • Kim, Jung-Hwan;Paek, Kyung-Soo;Kang, Wee-Kyung
    • Bulletin of the Korean Chemical Society
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    • 제23권2호
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    • pp.315-319
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    • 2002
  • Visible surface-assisted desorption/ionization mass spectrometry (SALDI-MS) has been investigated for several small macromolecules deposited on the graphite plate using laser radiation at 532 nm where most of the macromolecules are transparent. The graphite surface functioned well as a photon absorbing material and an energy transfer mediator for visible light. The results show that visible SALDI is a much softer ionization technique than UV-MALDI and FAB-MS in our results with synthetic macromolecules, PPG, PPGMBE and cavitand molecules. For the SALDI of biomolecules, glycerol as a proton source was essential with the graphite plate. As in visible SALDI, the role division of the photon absorbing material and the cationization agent can provide a generality in mass spectrometric analysis of macromolecules compared with MALDI using the dual functional matrix.

Optimization in Detecting Multiply-charged Protein Ions using MALDI TOF-MS

  • Lee, Jihyeon;Hong, Jangmi;Kim, Taehee;Kim, Jeongkwon
    • Mass Spectrometry Letters
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    • 제4권1호
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    • pp.21-23
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    • 2013
  • The effects of trifluoroacetic acid (TFA) were evaluated on the generation of multiply charged ions of cytochrome c in a 2-nitrophloroglucinol (2-NPG) matrix in high-vacuum, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The presence of 1% TFA in the 2-NPG matrix solution was more effective in generating multiply charged protein ions than matrix solutions containing 0.1% or 0% TFA. Regarding the matrix itself, with 1% TFA, 2-NPG was significantly more effective in generating multiply charged ions than 2,5-dihydroxybenzoic acid (2,5-DHB). The maximum charge state of cytochrome c was +8 when using a 2-NPG matrix containing 1% TFA.

Enhancement of Analyte Ionization in Desoprtion/Ionization on Porous Silicon (DIOS)-Mass Spectrometry(MS)

  • Lee Chang-Soo;Kim Eun-Mi;Lee Sang-Ho;KIm Min-Soo;Kim Yong-Kweon;Kim Byug-Gee
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제10권3호
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    • pp.212-217
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    • 2005
  • Desorption/ionization on silicon mass spectrometry (DIOS-MS) is a relatively new laser desorption/ionization technique for mass spectrometry without employing an organic matrix. This present study was carried to survey the experimental factors to improve the efficiency of DIOS-MS through electrochemical etching condition in structure and morphological properties of the porous silicon. The porous structure of silicon structure and its properties are crucial for the better performance of DIOS-MS and they can be controlled by the suitable selection of electrochemical conditions. The fabrication of porous silicon and ion signals on DIOS-MS were examined as a function of silicon orientation, etching time, etchant, current flux, irradiation, pore size, and pore depth. We have also examined the effect of pre- and post-etching conditions for their effect on DIOS-MS. Finally, we could optimize the electrochemical conditions for the efficient performance of DIOS-MS in the analysis of small molecule such as amino acid, drug and peptides without any unknown noise or fragmentation.

Proteome Analysis of Waito-c Rice Seedlings Treated with Culture Fluid of Gibberellin-producing Fungus, Fusarium proliferatum KGL0401

  • Rim, Soon-Ok;Lee, Jin-Hyung;Hwang, Seon-Kap;Suh, Seok-Jong;Lee, Jin-Man;Rhee, In-Koo;Kim, Jong-Guk
    • Journal of Microbiology and Biotechnology
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    • 제16권12호
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    • pp.1990-1994
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    • 2006
  • Fusarium proliferatum KGL0401 was previously isolated from Physalis alkekengi var. francheti plant roots and exhibited a high GA productivity. A gas chromatography-mass spectrometry (GC-MS) analysis of extracts of the culture fluid of F proliferatum KGL0401 also revealed the presence of $GA_1$, $GA_3$, $GA_4$, $GA_7$, $GA_{20}$, and $GA_{24}$. Therefore, the present study conducted a proteome analysis of waito-c rice treated with the culture fluid of the isolated F proliferatum KGL0401 to identify the protein expression triggered by the GA-containing culture fluid. The results revealed the overexpression of 180 protein spots in the sample treated with the culture fluid. Among them, 75 induced proteins were selected and analyzed by MALDI-TOF (matrix-assisted laser desorption-iorrization time-of-flight) mass spectrometry, followed by database searching, and 51 proteins were identified.

Free Radical Initiated Peptide Sequencing Using MALDI-TOF/TOF Mass Spectrometry

  • Song, Insu;Lee, Jae-ung;Baek, Jaehyeon;Cha, Sangwon;Han, Sang Yun;Oh, Han Bin
    • Mass Spectrometry Letters
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    • 제9권2호
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    • pp.56-60
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    • 2018
  • In this study, matrix-assisted laser desorption/ionization (MALDI) was applied to the TEMPO-assisted FRIPS for the first time. We found that 3-HPA is the optimal matrix for the analysis of p-TEMPO-Bz-Sc-peptides, which gives minimal precursor fragmentations. MALDI-TOF/TOF experiments on p-TEMPO-Bz-Sc-peptides yielded mainly $[a_n+H]^+$, $[z_n+H]^+$, and $[y_n]^+-type$ products, indicating that radical-driven peptide fragmentation occurs in MALDI-TOF/TOF-MS.