• 제목/요약/키워드: MALDI- TOF

검색결과 468건 처리시간 0.028초

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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    • 제11권5호
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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.

High-Throughput Screening Technique for Microbiome using MALDI-TOF Mass Spectrometry: A Review

  • Mojumdar, Abhik;Yoo, Hee-Jin;Kim, Duck-Hyun;Cho, Kun
    • Mass Spectrometry Letters
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    • 제13권4호
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    • pp.106-114
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    • 2022
  • A rapid and reliable approach to the identification of microorganisms is a critical requirement for large-scale culturomics analysis. MALDI-TOF MS is a suitable technique that can be a better alternative to conventional biochemical and gene sequencing methods as it is economical both in terms of cost and labor. In this review, the applications of MALDI-TOF MS for the comprehensive identification of microorganisms and bacterial strain typing for culturomics-based approaches for various environmental studies including bioremediation, plant sciences, agriculture and food microbiology have been widely explored. However, the restriction of this technique is attributed to insufficient coverage of the mass spectral database. To improve the applications of this technique for the identification of novel isolates, the spectral database should be updated with the peptide mass fingerprint (PMF) of type strains with not only microbes with clinical relevance but also from various environmental sources. Further, the development of enhanced sample processing methods and new algorithms for automation and de-replication of isolates will increase its application in microbial ecology studies.

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

  • 박정원;차재영;송재영;김희규;김범규;전병삼
    • 생명과학회지
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    • 제15권3호
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    • pp.427-433
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    • 2005
  • 초파리는 유전학, 발달 생물학, 행동 유전학, 노화 연구에 이르기까지 수많은 연구에 이용 되어져 왔다. 최근 초파리 전체 유전자의 염기서열이 보고되었으며, 유전자의 기능 분석과 발현 단백질에 대한 연구가 진행되고 있다. 본 연구에서는 야생형 초파리에서 추출한 단백질을 이차원 전기영동을 통해 400여개 이상의 단백질 스폿으로 분리하였으며, 각 스폿을 적출하여 트립신으로 처리하여 얻어진 펩타이드 단편을 MALDI-TOF-MS를 이용한 펩타이드 지문 검색으로 질량을 측정하였다. 측정된 질량을 초파리 데이터 베이스를 이용하여 분리한 단백질을 동정함으로써 59개의 유전자에서 발현되는 65개의 단백질 스폿을 동정하였다. 이러한 결과는 향후의 발생단계, 외부 자극, 노화 등에 관련되는 특이적 단백질 연구의 기초 자료로 활용될 수 있을 것이다.

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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    • 제27권9호
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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.

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
    • 대한의생명과학회지
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    • 제24권2호
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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.

홍합 [Mytilus coruscus]으로부터 항균활성 펩타이드의 정제 (Purification of a Antimicrobial Peptide from the Marine Mussel, Mytilus coruscus)

  • 김인혜;김진욱;이재화
    • 한국환경성돌연변이발암원학회지
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    • 제26권1호
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    • pp.25-29
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    • 2006
  • 흥합 (Mytilus coruscus) 추출물 RM 60을 사용하여 E. coli D31을 대상으로 항균활성 펩타이드를 분리 정제하였다. 정제한 펩타이드는 MALDI-TOF Mass spectrometry를 사용하여 분자량을 측정한 결과, 1464.92 Da로 나타났으며 Edman 분해법을 이용한 peptide sequencer를 사용하여 일차구조를 분석 중에 있다.

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Peptide C-terminal Sequence Analysis by MALDI-TOF MS Utilizing EDC Coupling with Br Signature

  • Shin, Man-Sup;Kim, Hie-Joon
    • Bulletin of the Korean Chemical Society
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    • 제32권4호
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    • pp.1183-1186
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    • 2011
  • The unique Br signature was utilized for C-terminal amino acid sequencing of model peptides. C-terminal carboxyl group was selectively derivatized in peptides, containing side chain carboxyl group, using 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC) and Br was introduced using 4-bromophenylhydrazine hydrochloride (BPH) in a one pot reaction. Matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) tandem mass spectra were obtained carrying the Br signature in the y-series ions. The Br signature facilitated C-terminal sequencing and discrimination of C-terminal carboxyl groups in the free acid and amide forms.

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.

Analysis of Branched PEG-Conjugated Interferon Alpha by Capillary Electrophoresis and MALDI- TOF Mass Spectrometry

  • Na, Dong-Hee;Park, Eun-Ji;Lee, Snag-Deuk;Jo, Young-Woo;Lee, Sung-Hee;Kim, Won-Bae;Lee, Kang-Choon
    • 대한약학회:학술대회논문집
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    • 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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    • pp.406.3-407
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    • 2002
  • Analysis of interferon alpha (IFN) modified with high molecular weight branched PEG was performed by capillary electrophoresis (CE) and MALDI-TOF mass spectrometry (MALDI-TOF MS). IFN was modified by the reaction of amine residues with an active ester of monomethoxy polyethylene glycol at various molar ratios. As a CE method. capilary electrophoresis sodium dodecyl sulfate nongel sieving (CE-SDS NGS) was performed using an uncoated capilary filled with a hydrophilic replaceable polymer network matrix. (omitted)

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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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    • 제35권10호
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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.