• Title/Summary/Keyword: MALDI-TOF-MS

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Identification of Salmonella spp. from porcine salmonellosis by matrix-assisted laser desorption ionization-time of flight mass spectrometry

  • Yang, Hyoung-Seok;Kim, Jae-Hoon
    • Korean Journal of Veterinary Service
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    • v.41 no.2
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    • pp.105-110
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    • 2018
  • A total of 41 Salmonella (S.) strains were isolated from pigs suffered with severe watery diarrhea and were tried to identify by both matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and polymerase chain reaction (PCR) analysis. Fibrinous exudate and ulceration in the large intestine were prevalent in gross observation, and variable degrees of enteritis were observed in the histology of large intestines. Subsequent polymerase chain reaction (PCR) analyses demonstrated that 41 strains were identified as S. Typhimurium (39 strains), though 2 stains were failed to identify. Further identification was performed using both direct smear and protein extraction method by MALDI-TOF MS analyses. In terms of extraction methods, 100% (41/41) of isolates were identified to species level of S. spp. Whereas only 43.9% (18/41) were identified to species level using the direct method. These results thus suggest that rapid and accurate diagnosis of porcine salmonellosis can be guaranteed by MALDI-TOF MS combined with protein extraction method.

Proteome Analysis of Drosophila melanogaster Used 2-DE and MALDI- TOF-MS (이차원 전기영동과 펩타이드 지문 검색법을 이용한 초파리의 프로테옴 분석)

  • Park Jeong-Won;Cha Jae-Young;Song Jae-Young;Kim Hee-Kyu;Kim Beom-Kyu;Jeon Beong-Sam
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.427-433
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    • 2005
  • With the completely discovery of the Drosophila genome sequence, the next great challenge is to extract its biological information by systematic expression and to perform functional analysis of the gene. Here we reported a proteome analysis of D. melanogaster with two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS). The cell extracts of D. melanogaster, $200{\mu}g$ were resolved to more than 400 silver-stained spots by 2-DE. The most abundant protein spots were ranged from 4.0-7.5 of pI and from 15-90 kDa of molecular weight. The excised spots were destained and in-gel digested by trypsin. The masses of the resulting peptide mixtures were measured by MALDI-TOF-MS. Identified proteins were compared with measured peptide mass and a dynamic peptide searching database which is accessible via the internet. The results revealed that identified proteins were produced by 59 genes derived from 65 protein spots.

Characterization of Synthetic Polyamides by MALDI-TOF Mass Spectrometry

  • Choi, Hae-Young;Choe, Eun-Kyung;Yang, Eun-Kyung;Jang, Sung-Woo;Park, Chan-Ryang
    • Bulletin of the Korean Chemical Society
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    • v.28 no.12
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    • pp.2354-2358
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    • 2007
  • MALDI-TOF-MS technique was applied to obtain structural and compositional information of synthetic polyamides, Nylon6 and Nylon66. Mass spectra of both the original and the hydrolyzed polyamide samples were analyzed using the self calibration method as well as the internal calibration method with the standard materials of known masses. The MALDI-TOF mass spectra of Nylon6 samples showed the presence of protonated, sodiated, and potassiated ions that were assigned to cyclic and NH2/COOH terminated linear oligomers. From the MALDI-TOF mass spectra of Nylon66 samples, the potassiated linear oligomers with three different end groups are identified as well as the cyclic oligomers, i.e., NH2/COOH terminated oligomers, NH2/NH2 terminated oligomers, and COOH/COOH terminated oligomers. Full characterization of the molecular species and end groups present in the polyamide samples has been achieved, and also the changes in mass spectral patterns after the hydrolysis of the samples are presented.

Availability of MADLDI-TOF MS for Identification of Gram Positive Bacilli Isolated from Blood Culture

  • Choi, Jin-Un;Kim, Sang-Ha;Hwang, Su-Jeong;Yu, Young-Bin;Kim, Sunghyun;Kim, Young-Kwon
    • Biomedical Science Letters
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    • v.24 no.2
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    • pp.108-115
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    • 2018
  • In the present study, results of the identification of Gram-positive bacilli (GPB) were analyzed by using the MALDI-TOF MS technique to score each 2-year blood culture at a university hospital. In addition, 16S rRNA sequence analyses and MALDI-TOF MS results are compared to targeting strains that had been isolated two or more times within the same patient, to evaluate the usefulness of MALDI-TOF MS in GPB identification. According to the cut-off (${\geq}1.7$) criteria, there were 410 (57.5%) reliable strains and 303 (42.5%) non-identified strains among the GPB identification results of 713 strains, using a microflex MALDI Biotyper (Bruker Daltonik GmbH, Bremen, Germany). The isolation appeared most often in the following order: Corynebacterium striatum, Bacillus cereus, Bacillus subtilis, Paenibacillus urinalis, and Listeria monocytogenes. Nearly three-fourths, 66 out of 89 (74.2%) of the strains for Corynebacterium striatum; 44 out of 60 (73.3%) strains for Bacillus cereus; and all (25 out of 25, 100%) Listeria monocytogenes strains were identified by their high scores of 2.0 or higher. Most (293 strains out of 303) non-identified strains were strains isolated only once and not significant as infectious bacilli. A total of 43 out of 50 (86.0%) strains matched and were able to be identified based on the 16 rRNA sequencing comparison results of strains that were isolated twice or more within the same patient and significant as infection bacilli. Non-matching among 5 out of 7 strains was not identified, even with MALDI-TOF MS. In conclusion, GPB can be identified in blood cultures using MALDI-TOF MS. This can be done accurately with ease, rapidly, and at a low cost. It is also thought to be helpful in GPB diagnosis and treatment.

Investigation of Angiotensin Glycosylation by MALDI-TOF and ESI Tandem Mass Spectrometry

  • Park, Soo-Jin;Park, Deok-Hie;Sul, Soo-Hwan;Oh, Sung-Hwan F.;Park, In-Sook;Chung, Doo-Soo;Kim, Hie-Joon;Kim, Min-Sik;Lee, Sang-Won
    • Bulletin of the Korean Chemical Society
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    • v.25 no.12
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    • pp.1791-1800
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    • 2004
  • Angiotensin I, a model decapeptide, was glycosylated and partially hydrolyzed with HCl (6 N, 80 $^{\circ}C$, 4 h), aminopeptidase, and carboxypeptidase Y. A single peptide mass map obtained from truncated peptides in the partial acid hydrolysate of angiotensin and its glycosylation product mixture by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry enabled sequencing of angiotensin by a combinatorial procedure. MALDI-TOF and electrospray ionization (ESI) tandem mass spectrometric results indicate that both the N-terminal amino group of aspartic acid and the guanidinium group of the second residue arginine are glycosylated.

Characterization of Poly(ethylene oxide)-b-Poly(L-lactide) Block Copolymer by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

  • Jeongmin Hong;Donghyun Cho;Taihyun Chang;Shim, Woo-Sun;Lee, Doo-Sung
    • Macromolecular Research
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    • v.11 no.5
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    • pp.341-346
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    • 2003
  • A poly(ethylene oxide)-b-poly(L-lactide) diblock copolymer (PEO-b-PLLA) is characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and a block length distribution map is constructed. Although the MALDI- TOF mass spectrum of PEO-b-PLLA is very complicated, most of the polymer species were identified by isolating the overlapped isotope patterns and by fitting the overlapped peaks to the Schulz-Zimm distribution function. Reconstructed MALDI-TOF MS spectrum was nearly identical to the measured spectrum and this method shows its potential to be developed as an easy and fast analysis method of low molecular weight block copolymers.

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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    • v.4 no.1
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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.

Selective Extraction and Quantification of Glutathione using Maleimide-Presenting Gold Nanoparticles

  • Oh, Hongseok;Lee, Jeongwook;Yeo, Woon-Seok
    • Bulletin of the Korean Chemical Society
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    • v.35 no.10
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    • pp.3047-3051
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    • 2014
  • In this paper, we describe a new method for the selective extraction and quantification of glutathione (GSH) using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and maleimide-presenting gold nanoparticles (Mal-AuNPs). Our strategy utilizes the Michael addition to selectively extract GSH, from chosen samples, onto the maleimide of Mal-AuNPs. After the extraction step, the GSH bound to the AuNPs was analyzed by MALDI-TOF MS in the presence of an internal standard which was prepared by reacting Mal-AuNPs with isotope-labeled GSH ($GSH^*$). The $GSH^*$ has the same structure as GSH but a higher molecular weight, and therefore, enables absolute quantification of GSH by comparing the mass signal intensities of the GSH- and $GSH^*$-conjugated alkanethiols. Our strategy was verified by analyzing GSH-spiked fetal bovine serum and NIH 3T3 cells.

Ultrasonic Monitoring of Reaction of Fullerene$[C_{60}]$ with 3-Chloroperoxy Benzoic acid (풀러렌$[C_{60}]$과 3- Chloroperoxy Benzoic acid 반응의 초음파적 조사)

  • Ko, Weon-Bae;Buckin, Vitaly
    • Elastomers and Composites
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    • v.41 no.1
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    • pp.57-62
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    • 2006
  • The reaction of fullerene$[C_{60}]$ and 3-chloroperoxy benzoic acid in 1,2-dichlorobenzene was monitored by high resolution ultrasonic spectroscopy and the product of reaction by fullerene $[C_{60}]$ and 3-chloroperoxy benzoic acid in 1,2-dichlorobenzene was characterized using MALDI-TOF-MS spectra.

Rapid Identification of Vibrio Species Isolated from the Southern Coastal Regions of Korea by MALDI-TOF Mass Spectrometry and Comparison of MALDI Sample Preparation Methods

  • Cho, Youngjae;Kim, Eiseul;Han, Sun-Kyung;Yang, Seung-Min;Kim, Mi-ju;Kim, Hyun-Joong;Kim, Chang-Gyeom;Choo, Dong-Won;Kim, Young-Rok;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1593-1601
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    • 2017
  • Vibrio species are generally recognized as pathogens predominant in seafood along coastal areas. The food industry has sought to develop efficient microbial detection methods. Owing to the limits of conventional methods, this study aimed to establish a rapid identification method for Vibrio isolated from Korea, based on matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Four different preparation procedures were compared to determine the appropriate means to pretreat Vibrio species, using 17 isolates and five reference strains. Extended direct transfer and full formic acid extraction methods using bacterial colonies on agar plates revealed very low identification rates. Formic acid and trifluoroacetic acid (TFA) extractions using bacterial broth cultures were also performed. All Vibrio isolates and reference strains prepared by TFA extraction were successfully identified to the species level (17/22, 77.3%) and to the genus level (5/22, 22.7%). Thus, TFA extraction was considered the most appropriate method to pretreat Vibrio species for MALDI-TOF MS. The remaining 33 isolates and two reference strains were prepared by TFA extraction and analyzed by MALDI-TOF MS. Overall, 50 isolates were identified to the species level (40/50, 80%) and to the genus level (10/50, 20%). All isolates were identified as 43 V. alginolyticus, six V. parahaemolyticus, and one V. vulnificus species. V. alginolyticus and V. parahaemolyticus were isolated from fish offal (87.5% and 12.5%, respectively), seawater (91.3%, 8.7%), and shellfish (62.5%, 37.5%), whereas V. alginolyticus and V. vulnificus were isolated from sediment (90.9% and 9.1%, respectively). This study established a reliable system of MALDI-TOF MS preparation and analysis for Vibrio identification.