• Title/Summary/Keyword: MALDI-TOF MS/MS

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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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    • v.32 no.4
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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.

Matrix-assisted Laser Desorption/ Ionization Time-of-flight Mass Spectrometry를 이용한 화장품에서의 계면활성제 분석

  • 이명희;김상진
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.25 no.3
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    • pp.1-21
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    • 1999
  • 화장품, 의료용, 가정용품 및 공업용제품에까지 광범위한 용도로 사용량이 많은 중요한 비이온 계면활성제의 종류 중에서 polyoxyethylene(POE) 부가형 계면활성제의 경우 그 부가몰수에 따라 특성이 달라지고 용도가 다르게 사용된다. 이때 부가된 분자들의 분자량이 크고 분포를 이루는 혼합물이기 때문에 분석이 매우 어렵다. 따라서 MALDI-TOF/MS 방법을 이용하여 이들의 분자량과 그 분포를 측정함으로써 쉽고 빠르게 측정할 수 있는 새로운 방법을 개발하고자 하였다. 이 논문에서는 화장품에 주로 사용되는 비이온 계면활성제를 선택하여 MALDI-TOF/AfS를 측정하여 스펙트럼으로부터 분자량 분포와 POE부가정도를 측정할 수 있었다. 그리고 이 조건을 적용하여 시중에 판매되는 제품에서 추출된 비이온 계면활성제의 MALDI-TOF/MS 스펙트럼으로부터 분자량 분포와 POE 부가 정도를 확인 할 수 있었다.

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Discrimination of Bacillus anthracis Spores by Direct in-situ Analysis of Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry

  • Jeong, Young-Su;Lee, Jonghee;Kim, Seong-Joo
    • Bulletin of the Korean Chemical Society
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    • v.34 no.9
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    • pp.2635-2639
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    • 2013
  • The rapid and accurate identification of biological agents is a critical step in the case of bio-terror and biological warfare attacks. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry has been widely used for the identification of microorganisms. In this study, we describe a method for the rapid and accurate discrimination of Bacillus anthracis spores using MALDI-TOF MS. Our direct in-situ analysis of MALDI-TOF MS does not involve subsequent high-resolution mass analyses and sample preparation steps. This method allowed the detection of species-specific biomarkers from each Bacillus spores. Especially, B. anthracis spores had specific biomarker peaks at 2503, 3089, 3376, 6684, 6698, 6753, and 6840 m/z. Cluster and PCA analyses of the mass spectra of Bacillus spores revealed distinctively separated clusters and within-groups similarity. Therefore, we believe that this method is effective in the real-time identification of biological warfare agents such as B. anthracis as well as other microorganisms in the field.

Preparation of Fullerene Oxides by Fullerenes[$C_{60},C_{70}$] with Several Oxidants under Ultrasonic Condition (초음파 조건에서 산화제를 이용한 풀러렌 산화물의 제조)

  • Kown, Sock-Chan;Jeong, Hong-Seok;Ko, Weon-Bae
    • Elastomers and Composites
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    • v.38 no.2
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    • pp.147-156
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    • 2003
  • Synthesis of fullerene oxides by fullerenes [$C_{60},\;C_{70}$] and several oxidants such as benzoylperoxide, trichloroisocyanuric acid, methyltrioxorhenium(VII), iodosobenzene, phosphorous pentoxide take place under ultrasonic condition at room temperature. The MALDI-TOF MS,UV-visible spectra and HPLC analysis confirmed that the products of fullerenes oxidation are [$C_{60}(O)_n$], ($n=1{\sim}3$ or n=1) and [$C_{70}(O)_n$], ($n=1{\sim}2$ or n=1). As compared with the reactivity of epoxidation of fullerenes [$C_{60},\;C_{70}$], the reaction rate of $C_{70}$ was lower than that of $C_{60}$ under same reaction condition.

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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    • v.13 no.4
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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.

Identification of Lactic Acid Bacteria in Galchi- and Myeolchi-Jeotgal by 16S rRNA Gene Sequencing, MALDI-TOF Mass Spectrometry, and PCR-DGGE

  • Lee, Yoonju;Cho, Youngjae;Kim, Eiseul;Kim, Hyun-Joong;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.7
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    • pp.1112-1121
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    • 2018
  • Jeotgal is a Korean traditional fermented seafood with a high concentration of salt. In this study, we isolated lactic acid bacteria (LAB) from galchi (Trichiurus lepturus, hairtail) and myeolchi (Engraulis japonicas, anchovy) jeotgal on MRS agar and MRS agar containing 5% NaCl (MRS agar+5% NaCl), and identified them by using 16S rRNA gene sequencing and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) as culture-dependent methods. We also performed polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) as a culture-independent method to identify bacterial communities. Five samples of galchi-jeotgal and seven samples of myeolchi-jeotgal were collected from different regions in Korea. A total of 327 and 395 colonies were isolated from the galchi- and myeolchi-jeotgal samples, respectively. 16S rRNA gene sequencing and MALDI-TOF MS revealed that the genus Pediococcus was predominant on MRS agar, and Tetragenococcus halophilus on MRS agar+5% NaCl. PCR-DGGE revealed that T. halophilus, Tetragenococcus muriaticus, and Lactobacillus sakei were predominant in both types of jeotgal. T. halophilus was detected in all samples. Even though the same species were identified by both culture-dependent and -independent methods, many species identified by the culture-dependent methods were not in the bacterial list identified by the culture-independent methods. The distribution of bacteria in galchi-jeotgal was more diverse than in myeolchi-jeotgal. The diverse LAB in galchi- and myeolchi-jeotgals can be further studied as candidates for starter cultures to produce fermented foods.

2-DE and MALDI-TOF MS-based identification of bovine whey proteins in milk collected soon after parturition

  • Lee, Jae Eun;Lin, Tao;Kang, Jung Won;Shin, Hyun Young;Lee, Joo Bin;Jin, Dong Il
    • Korean Journal of Agricultural Science
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    • v.45 no.4
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    • pp.635-643
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    • 2018
  • Bovine milk is widely consumed by humans and is a primary ingredient of dairy foods. Proteomic approaches have the potential to elucidate complex milk proteins and have been used to study milk of various species. Here, we performed a proteomic analysis using 2-dimensional electrophoresis (2-DE) and matrix assisted laser desorption ionization-time of flight mass spectrometer (MALDI-TOF MS) to identify whey proteins in bovine milk obtained soon after parturition (bovine early milk). The major casein proteins were removed, and the whey proteins were analyzed with 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE). The whey proteins (2 mg) were separated by pI and molecular weight across pH ranges of 3.0 - 10.0 and 4.0 - 7.0. The 2-DE gels held about 300 to 700 detectable protein spots. We randomly picked 12 and nine spots that were consistently expressed in the pH 3.0 - 10.0 and pH 4.0 - 7.0 ranges, respectively. Following MALDI-TOF MS analysis, the 21 randomly selected proteins included proteins known to be present in bovine milk, such as albumin, lactoferrin, serum albumin precursor, T cell receptor, polymeric immunoglobulin receptor, pancreatic trypsin inhibitor, aldehyde oxidase and microglobulin. These proteins have major functions in immune responses, metabolism and protein binding. In summary, we herein identified both known and novel whey proteins present in bovine early milk, and our sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed their expression pattern.

Multidrug-resistant Staphylococci Found on Book Surfaces in East London Libraries

  • Idris, Adi;Cutler, Ron R
    • Microbiology and Biotechnology Letters
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    • v.45 no.4
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    • pp.354-357
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    • 2017
  • There is an increase in the presence of drug-resistant staphylococci outside of the nosocomial and healthcare setting. Although the presence of staphylococci has been studied in several public spaces, nothing is known on the presence of staphylococci in public libraries. Book surfaces from public libraries in the East London area, United Kingdom were swabbed and cultured and identity of the isolates determined by matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS). Seven different staphylococcal species were identified by MALDI-TOF-MS analysis. This short study provides evidence of the presence of multidrug-resistant staphylococci in public libraries in the East London area.

Evaluation of MALDI Biotyping for Rapid Subspecies Identification of Carbapenemase-Producing Bacteria via Protein Profiling

  • Somboro, Anou M.;Tiwari, Dileep;Shobo, Adeola;Bester, Linda A.;Kruger, Hendrik G.;Govender, Thavendran;Essack, Sabiha Y.
    • Mass Spectrometry Letters
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    • v.5 no.4
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    • pp.110-114
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    • 2014
  • The method of direct mass spectrometry profiling is reliable and reproducible for the rapid identification of clinical isolates of bacteria and fungi. This is the first study evaluating the approach of MALDI-TOF mass spectrometry profiling for rapid identification of carbapenemase-resistant enterobacteriaceae (CRE). Proof of concept was achieved by the discrimination of CRE using MALDI Biotyper MS based on the protein. This profiling appears promising by the visual observation of consistent unique peaks, albeit low intensity, that could be picked up from the mean spectra (MSP) method. The Biotyper MSP creation and identification methods needed to be optimized to provide significantly improved differences in scores to allow for subspecies identification with and without carbapenemases. These spectra were subjected to visual peak picking and in all cases; there were pertinent differences in the presence or absence of potential biomarker peaks to differentiate isolates. We also evaluated this method for potential discrimination between different carbapenemases bacteria, utilizing the same strategy. Based on our data and pending further investigation in other CREs, MALDI-TOF MS has potential as a diagnostic tool for the rapid identification of even closely related carbapenemases but would require a paradigm shift in which Biotyper suppliers enable more flexible software control of mass spectral profiling methods.

Comparative proteomic analysis of plant responses to sound waves in Arabidopsis

  • Kwon, Young Sang;Jeong, Mi-Jeong;Cha, Jaeyul;Jeong, Sung Woo;Park, Soo-Chul;Shin, Sung Chul;Chung, Woo Sik;Bae, Hanhong;Bae, Dong-Won
    • Journal of Plant Biotechnology
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    • v.39 no.4
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    • pp.261-272
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    • 2012
  • Environmental factors greatly influence the growth, development, and even genetic characteristics of plants. The mechanisms by which sound influences plant growth, however, remain obscure. Previously, our group reported that several genes were differentially regulated by specific frequenciesof sound treatmentusing a sound-treated subtractive library. In this study, we used a proteomic approach to investigate plant responses to sound waves in Arabidopsis. The plants were exposed to 250-Hz or 500-Hz sound waves, and total proteins were extracted from leaves 8 h and 24 h after treatment. Proteins extracted from leaves were subjected to 2-DE analysis. Thirty-eight spots were found to be differentially regulated in response to sound waves and were identified using MALDI-TOF MS and MALDI-TOF/TOF MS. The functions of the identified proteins were classified into photosynthesis, stress and defense, nitrogen metabolism, and carbohydrate metabolism. To the best of our knowledge, this is the first report on the analysis of protein changes in response to sound waves in Arabidopsis leaves. These findings provide a better understanding of the molecular basis of responses to sound waves in Arabidopsis.