• Title/Summary/Keyword: MALDI-TOF Mass

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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.

Simple measurement the chelator number of antibody conjugates by MALDI-TOF MS

  • Shin, Eunbi;Lee, Ji Woong;Lee, Kyo Chul;Shim, Jae Hoon;Cha, Sangwon;Kim, Jung Young
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.3 no.2
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    • pp.54-58
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    • 2017
  • Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS) is one of the powerful methods that enable analysis of small molecules as well as large molecules up to about 500,000 Da without severe fragmentation. MALDI-TOF MS, thus, has been a very useful an analytical tool for the confirmation of synthetic molecules, probing PTMs, and identifying structures of a given protein. In recent nuclear medicine, MALDI-TOF MS liner ion mode helps researcher calculate the average number of chelator(or linkage) per an antibody conjugate, such as DOTA-(or DFO-) trastuzumab for labeling a medical radioisotope. This simple technique can be utilized to improve the labeling method and control the quality at the development of antibody-based radiopharmaceuticals, which is very effected to diagnosis and therapy for in vivo tumor cells, with radioisotopes like $^{89}Zr$, $^{64}Cu$, and 177Lu. To minimize the error, MALDI-TOF MS measurement is repeatedly performed for each sample in this study, and external calibration is carried out after data collection.

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.

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

  • Kim In-Hae;Kim Jin-Wook;Lee Jae-Hwa
    • Environmental Mutagens and Carcinogens
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    • v.26 no.1
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    • pp.25-29
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    • 2006
  • Antimicrobial peptides (AMPs) play an important role in this response by rapidly killing invading microorganisms. In this study antimicrobial peptide has been isolated from acidified whole body extract of a bivalve mollusk, the marine mussel (Mytilus coruscus). This peptide purified to homogeneity by gel-filtration and reversed-phase high performance liquid chromatography. The molecular weight was 1464.92 Da, determined by MALDI-TOF Mass spectrometry. In addition to growth inhibition of Escherichia coli D31.

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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.

MALDI-TOF Mass Spectrometry as a Useful Tool for Identification of Enterococcus spp. from Wild Birds and Differentiation of Closely Related Species

  • Stepien-Pysniak, Dagmara;Hauschild, Tomasz;Rozanski, Pawel;Marek, Agnieszka
    • Journal of Microbiology and Biotechnology
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    • v.27 no.6
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    • pp.1128-1137
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    • 2017
  • The aim of this study was to explore the accuracy and feasibility of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) in identifying bacteria from environmental sources, as compared with rpoA gene sequencing, and to evaluate the occurrence of bacteria of the genus Enterococcus in wild birds. In addition, a phyloproteomic analysis of certain Enterococcus species with spectral relationships was performed. The enterococci were isolated from 25 species of wild birds in central Europe (Poland). Proteomic (MALDI-TOF MS) and genomic (rpoA gene sequencing) methods were used to identify all the isolates. Using MALDI-TOF MS, all 54 (100%) isolates were identified as Enterococcus spp. Among these, 51 (94.4%) isolates were identified to the species level (log(score) ${\geq}2.0$), and three isolates (5.6%) were identified at a level of probable genus identification (log(score) 1.88-1.927). Phylogenetic analysis based on rpoA sequences confirmed that all enterococci had been correctly identified. Enterococcus faecalis was the most prevalent enterococcal species (50%) and Enterococcus faecium (33.3%) the second most frequent species, followed by Enterococcus hirae (9.3%), Enterococcus durans (3.7%), and Enterococcus casseliflavus (3.7%). The phyloproteomic analysis of the spectral profiles of the isolates showed that MALDI-TOF MS is able to differentiate among similar species of the genus Enterococcus.

Rapid Identification of Staphylococcus Species Isolated from Food Samples by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

  • Kim, Eiseul;Kim, Hyun-Joong;Yang, Seung-Min;Kim, Chang-Gyeom;Choo, Dong-Won;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.29 no.4
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    • pp.548-557
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    • 2019
  • Staphylococcus species have a ubiquitous habitat in a wide range of foods, thus the ability to identify staphylococci at the species level is critical in the food industry. In this study, we performed rapid identification of Staphylococcus species using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight mass spectrometry (MALDI-TOF MS). MALDI-TOF MS was evaluated for the identification of Staphylococcus reference strains (n = 19) and isolates (n = 96) from various foods with consideration for the impact of sample preparation methods and incubation period. Additionally, the spectra of isolated Staphylococcus strains were analyzed using principal component analysis (PCA) and a main spectra profile (MSP)-based dendrogram. MALDI-TOF MS accurately identified Staphylococcus reference strains and isolated strains: the highest performance was by the EX method (83.3~89.5% accuracy) at species level identification (EDT, 70.3~78.9% accuracy; DT, less than 46.3~63.2% accuracy) of 24-h cultured colonies. Identification results at the genus level were 100% accurate at EDT, EX sample preparation and 24-h incubation time. On the other hand, the DT method showed relatively low identification accuracy in all extraction methods and incubation times. The analyzed spectra and MSP-based dendrogram showed that the isolated Staphylococcus strains were characterized at the species level. The performance analysis of MALDI-TOF MS shows the method has the potential ability to discriminate between Staphylococcus species from foods in Korea. This study provides valuable information that MALDI-TOF MS can be applied to monitor microbial populations and pathogenic bacteria in the food industry thereby contributing to food safety.

MALDI-TOF MS System for the Identification of Microorganisms in Milk and Dairy Products (우유 및 유제품 중 미생물 동정을 위한 MALDI-TOFMS활용)

  • Kim, Hyoun Wook;Ham, Jun-Sang;Seol, Kuk-Hwan;Han, Sangha;Park, Beam Young;Oh, Mi-Hwa
    • Journal of Dairy Science and Biotechnology
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    • v.30 no.2
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    • pp.131-137
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    • 2012
  • Rapid and reliable identification of microorganisms is a key for tracing the relationship between the target bacteria and related infectious diseases. Various identification methods such as classical phenotypic analysis, numerical taxonomic analysis, and DNA sequencing have been widely used to classify microorganisms in milk and dairy products. Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) identifies targeted bacteria in milk and milk products. Several studies have demonstrated that MALDI-TOF MS identification is an efficient and inexpensive method for the rapid and routine identification of isolated bacteria. MALDI-TOF MS could provide accurate identification of bacteria in milk and milk products at the serotype or strain level and enable antibiotic resistance profiling within minutes.

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Development of a Rapid and Accurate Identification Method for Citrobacter Species Isolated from Pork Products Using a Matrix-Assisted Laser-Desorption Ionization Time-of-Flight Mass Spectrometry (MALDITOF MS)

  • Kwak, Hye-Lim;Han, Sun-Kyung;Park, Sunghoon;Park, Si Hong;Shim, Jae-Yong;Oh, Mihwa;Ricke, Steven C.;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.25 no.9
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    • pp.1537-1541
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    • 2015
  • Previous detection methods for Citrobacter are considered time consuming and laborious. In this study, we have developed a rapid and accurate detection method for Citrobacter species in pork products, using matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). A total of 35 Citrobacter strains were isolated from 30 pork products and identified by both MALDI-TOF MS and 16S rRNA gene sequencing approaches. All isolates were identified to the species level by the MALDI-TOF MS, while 16S rRNA gene sequencing results could not discriminate them clearly. These results confirmed that MALDITOF MS is a more accurate and rapid detection method for the identification of Citrobacter species.

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.